Biomedical subjects

M L Day

Publications and source records attributed to M L Day.

At least 55 records · Page 3Linked to original sources

PubMed1998

Cell cycle regulation of a T-type calcium current in early mouse embryos.

In this paper we report that the amplitude of the T-current changes in a cell cycle-dependent manner in single- and two-cell embryos, being large in unfertilized oocytes and decreasing after fertilization throughout the first cell cycle to a minimum in early M phase, before increasing again during late telophase. This increase can be prevented by arresting the cell cycle in metaphase, but is not prevented by cytochalasin D. During the second cell cycle the T-current remains large until late G2 phase when it decreases. Protein synthesis inhibition has no effect on the changes in T-current amplitude during the cell cycle. This finding, along with the observation that the size of the T-current does not increase until mitosis exit, suggests that cell cycle regulation of the current does not involve the cycling of cyclin-dependent kinase (cdk)1/cyclin B activity. Inhibition of the T-current with pimozide has no effect on cleavage. The cell cycle-dependent changes in T-current amplitude appear to be driven by the same cdk1/cyclin-B-independent clock that we have previously shown to drive cyclic changes in K+ channel activity in early mouse embryos. Thus, multiple ion transport systems in the preimplantation mouse embryo are controlled by this cdk1/cyclin-B-independent clock.

Animals↗

PubMed1998

Changes in ovarian follicles following acute infection with bovine viral diarrhea virus.

Bovine viral diarrhea virus (BVDV) has been associated with several reproductive problems in cattle, including poor fertility, early embryonic deaths, abortion and congenital anomalies. Little is known about the cause of poor fertility in cows acutely infected with BVDV. The purpose of this study was to identify changes in ovarian function following acute infection with noncytopathic BVDV. The ovaries of 5 BVDV sero-negative and virus-negative pubertal heifers were monitored daily for 4 consecutive estrous cycles. The position and diameter of all follicles (> 5 mm) and luteal structures were recorded. Daily plasma samples were collected to measure peripheral progesterone and estradiol levels. Each heifer was infected intranasally with noncytopathic BVDV following ovulation of the second estrous cycle. The maximum diameter and growth rate of dominant anovulatory and ovulatory follicles were significantly reduced following acute BVDV infection. Similarly, the number of subordinate follicles associated with both the anovulatory and ovulatory follicle was reduced following infection. There were no significant differences in other follicle or luteal dynamic parameters or in peripheral progesterone or estradiol levels. Ovarian follicular growth was different during the first 2 estrous cycles following acute infection with BVDV when compared with the 2 estrous cycles preceding infection. These differences may be important in explaining reduced fertility in herds with acute BVDV infection.

Acute Disease↗

PubMed1998

Duration of treatment with progesterone and regression of persistent ovarian follicles in cattle.

The objective of the present study was to determine the duration of elevated concentrations of progesterone necessary to induce atresia of persistent ovarian follicles. Heifers were administered 25 mg of PGF2alpha on d 6 and 7 (d 0 = d of synchronized estrus) and a norgestomet implant from d 6 to 14. Ovaries were monitored by ultrasonography, and blood samples were collected on d 3, 5, 7, 9, 11, and 12 and daily from d 14 until ovulation. On d 12, heifers received either two progesterone-releasing intravaginal devices (PRID) for 6 h (6-h; n = 5), two PRID for 24 h (24-h; n = 5), or no treatment (CON; n = 5). Blood samples were collected at 15-min intervals from h -6 to 30 (PRID insertion = h 0) and analyzed for concentrations of LH. Characteristics of LH secretion were determined for consecutive 6-h periods (Period 0 to 5). Hourly blood samples, collected from h 0 to 29, were analyzed for concentrations of 17beta-estradiol (estradiol) and progesterone. The dominant ovarian follicles present on d 7 increased in size to 15.4+/-.3 mm on d 12 ("persistent follicle"). Following removal of the PRID and norgestomet implants, atresia of persistent follicles and ovulation of new follicles were induced in one of five and in four of five heifers in the 6-h and 24-h treatments, respectively. Persistent follicles ovulated after withdrawal of norgestomet in all other heifers. Concentrations of progesterone were increased from h 1 to 7 in the 6-h and h 1 to 26 in the 24-h treatment. Frequency of LH pulses was reduced (P < .05) during Periods 1 to 2 in the 6-h and Periods 1 to 5 in the 24-h treatment relative to the CON treatment. By h 10, concentrations of estradiol in the 6-h and 24-h treatments were lower (P < . 05) than in the CON treatment. This suppression continued through h 29 in the 24-h treatment (P < .05), whereas concentrations in the 6-h treatment were intermediate to those of the CON and 24-h treatments after h 14. Suppression of pulsatile LH release and estradiol secretion was evident with 6 and 24 h of treatment with progesterone, but only the 24-h treatment induced atresia of persistent follicles in a majority of the heifers.

Administration, Intravaginal↗

PubMed1998

Development of a progestin-based estrus synchronization program: I. Reproductive response of cows fed melengestrol acetate for 20 days with an injection of progesterone.

We designed two experiments to determine the efficacy of an estrus control system in cows that combined long-term progestin exposure (20 d) with an acute increase in progesterone concentration. In Exp. 1, cows (n = 30) were fed either melengestrol acetate (MGA; .5 mg x cow(-1) x d(-1)) or ground ear corn (MGA carrier) for 20 d. On d -15 (last day of MGA feeding = d 0), cows were administered 25 mg of PGF2alpha to regress the corpus luteum (CL) and establish an environment conducive to the development of persistent follicles. To synchronously regress persistent follicles, cows fed MGA (n = 15) were injected with 200 mg of progesterone on d -2 (MGA-P), and the cows fed the MGA carrier were not treated (CONT; n = 15). Cows in the CONT group were artificially inseminated 12 h after detection of spontaneous estrus from d -20 to d 8. Estrus was observed, and all cows in the MGA-P group were artificially inseminated during the period of estrus synchronization (SYNC; d 1 to 8). No difference in conception rate was observed between treatments. In Exp. 2, postpartum cows (n = 113) received either the MGA-P (n = 56) or CONT (n = 57) treatment. More (P < .05) cows were observed in estrus during SYNC in the MGA-P (50%) than in the CONT (28%) group. Of the cows in the MGA-P group that were not observed in estrus during SYNC, 50% were in estrus for the first time 23 to 29 d after MGA withdrawal (SYNC2), suggesting that these cows ovulated without observable estrus during SYNC. Estrus was observed for the first time during SYNC2 in more (P < .05) cows in the MGA-P (25%) than in the CONT (7%) group. Conception rate at the synchronized estrus, pregnancy rate, and interval to first service and pregnancy were similar between treatments. We conclude that administration of MGA-P results in the synchronization and(or) induction of a fertile estrus in cows.

Administration, Oral↗

PubMed1998

Development of a progestin-based estrus synchronization program: II. Reproductive response of cows fed melengestrol acetate for 14 days with injections of progesterone and prostaglandin F2alpha.

We tested the efficacy of an estrus control system designed to provide optimal control of follicular development. In Exp. 1, postpartum cows (n = 133) and yearling heifers (n = 57) were fed either .5 mg x female(-1) x d(-1) of melengestrol acetate (MGA) or the carrier for MGA from d -13 to d 0 (d 0 = last day of MGA feeding). All females received 25 mg of PGF2alpha (i.m.) on d -13 and 0. On d -6, cows and heifers fed MGA were administered an i.m. injection of progesterone (200 mg; MGA/P4), and those fed the corn carrier (2XPGF2alpha) received no progesterone. Beginning on d 1, females were bred by AI from d 1 to at least d 5. During the estrus synchronization period (d 1 to d 5), more (P < .05) postpartum cows were observed in estrus (70.1 vs 42.4%), the timing of estrus was more (P < .05) precise, conception rate was similar, and pregnancy rate was higher (P < .05) in the MGA/P4 than in the 2XPGF2alpha treatment. More (P < .05) cows that were anestrous at the beginning of the breeding season were in estrus during the synchronization period in the MGA/P4 (55.8%) than in the 2XPGF2alpha (28.6%) treatment. In heifers, estrus was synchronized in over 90% of females, and neither conception nor pregnancy rate during the synchronization period differed between treatments. In Exp. 2, postpartum cows (n = 122) and heifers (n = 84) received treatments (MGA/P4 or 2XPGF2alpha) as described for Exp. 1 with one exception. In the MGA/ P4 treatment, progesterone was administered on d -7 rather than d -6. Females were bred by AI from d 1 to 5. The estrus response and conception rate during the synchronization period did not differ between treatments for either cows or heifers. We conclude that the progestin-based estrous synchronization system used in this study effectively synchronized an estrus of normal fertility in cyclic cows and induced a majority of anestrous cows to reinitiate estrous cycles.

Administration, Oral↗

PubMed1997

Retinoblastoma protein-dependent growth signal conflict and caspase activity are required for protein kinase C-signaled apoptosis of prostate epithelial cells.

Both protein kinase C and the retinoblastoma tumor suppressor protein have been linked to the regulation of cell growth and cell death, suggesting the differential roles these factors play in mediating cell fate. In some cells, protein kinase C-induced activation of the retinoblastoma protein results in G1 arrest. However, inducible overexpression and activation of the protein kinase Calpha isozyme or the addition of 12-O-tetradecanoylphorbol-13-acetate in the prostate epithelial cell line, LNCaP, resulted in apoptosis preceded by induction of p21 and dephosphorylation of the retinoblastoma protein. Consistent with a role for the retinoblastoma growth suppressor protein in protein kinase C-induced apoptosis, DU145 cells, which do not express functional retinoblastoma protein or LNCaP cells, which have been transfected with the retinoblastoma inhibitor, E1a, were resistant to apoptosis. LNCaP apoptosis was initiated by a unique conflict between the growth-suppressive activity of the retinoblastoma protein and growth-promoting mitogenic signals. Thus, when this conflict was prevented by serum depletion, apoptosis was suppressed. The caspase family of cysteine proteases is believed to encompass the execution machinery of mammalian apoptosis, and addition of the cell-permeable caspase inhibitor, Z-Val-Ala-Asp-fluoromethylketone, afforded nearly total protection from protein kinase C-signaled apoptosis. This protection correlated with the total loss of caspase activity as measured by the proteolytic cleavage of nuclear poly(ADP-ribose) polymerase. On the basis of these results, we propose that protein kinase C regulates a novel cell death pathway that is initiated by a cellular conflict between retinoblastoma growth-suppressive signals and serum mitogenic signals in proliferating prostate epithelial cells and that is executed by the caspase family of cysteine proteases.

Adenovirus E1A Proteins↗

PubMed1997

Cell anchorage regulates apoptosis through the retinoblastoma tumor suppressor/E2F pathway.

Epithelial cells are dependent upon adhesion to extracellular matrix for survival. We show that loss of beta1 integrin receptor contact with extracellular matrix signals the inhibition of G1 cyclin-dependent kinase activity. This loss of cyclin-dependent kinase activity leads to accumulation of the hypophosphorylated (active) form of the retinoblastoma tumor suppressor protein (Rb). We present evidence that in epithelial cells deprived of matrix contact, the growth suppression signal elicited by hypophosphorylated Rb opposes stimulatory signals from serum growth factors, leading to a cell cycle conflict that triggers apoptosis. This apoptotic pathway is modulated by Bcl-2 through a novel mechanism that regulates Rb phosphorylation. We present evidence that the Rb-dependent apoptotic pathway functions in vivo in the apoptosis of the prostate glandular epithelium following castration.

Animals↗

PubMed1997

Control of cytosolic pH in two-cell mouse embryos: roles of H(+)-lactate cotransport and Na+/H+ exchange.

In this study we used imaging techniques with the fluorescent pH-sensitive dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein to investigate the control of cytosolic pH (pHi) in two-cell mouse embryos in nominally HCO3(-)-free conditions. We found that the resting pHi of two-cell embryos (40-50 h after human chorionic gonadotropin) in HCO3(-)-free M2 was 7.31 +/- 0.01 (n = 172 embryos), which is significantly above the level predicted if H+ is at electrochemical equilibrium. We showed that two-cell embryos contain a H(+)-monocarboxylate cotransport system with apparent Michaelis constants for D-lactate, L-lactate, and pyruvate of 11.5, 3.7, and 3.5 mM, respectively. It is inhibited by p-chloromercuribenzoic acid (300 microM), p-chloromercuriphenylsulfonic acid (300 microM), and alpha-cyano-4-hydroxycinnamate (1 mM) and is insensitive to 4,4'-diisothiocyanodihydrostilbene-2,2'-disulfonic acid (500 microM). We also showed that the pHi response to the acid load produced by an NH4Cl pulse has two components, one due to H(+)-monocarboxylate cotransport and the other due to Na+/H+ exchange. We found no evidence that a H+ conductance was responsible in these cells for the recovery in pHi after an acid load.

Ammonium Chloride↗

PubMed1997

Estrus and luteal function in suckled beef cows that were anestrous when treated with an intravaginal device containing progesterone with or without a subsequent injection of estradiol benzoate.

The objectives in this study were to determine whether treatment with progesterone (P4) via an intravaginal device would induce estrus and development of corpora lutea (CL) with typical life spans and whether treatment with estradiol benzoate (EB) following device removal would enhance the responses. At treatment initiation (d 0), suckled beef cows (n = 362) that were anestrous received one of the following: 1) an intravaginal device containing P4 for 7 d plus an injection of 1 mg of EB 24 to 30 h after device removal (P4+EB), 2) an intravaginal device containing P4 for 7 d (P4), 3) a sham device for 7 d plus an injection of 1 mg of EB 24 to 30 h after device removal (EB), or 4) a sham device for 7 d (control). Progesterone treatment increased the proportion of cows that formed CL with a typical lifespan (P < .001) and exhibited behavioral estrus (P < .05). Treatment with EB in combination with P4 increased (P < .001) the proportion of cows that exhibited estrus. Treatment with P4 alone had no effect, but the combination of P4 and EB increased (P < .01) the proportion of cows that formed short-lived or typical lifespan CL by the end of the experiment compared with untreated cows. In summary, treatment with P4 increased the incidence of estrus and the proportion of cows that formed CL with a typical lifespan. Treatment with P4 resulted in resumption of luteal function in suckled beef cows that were anestrous, with most cows developing CL with a typical lifespan, whereas EB enhanced the expression of estrus.

Administration, Intravaginal↗

PubMed1996

Progestin-induced puberty and secretion of luteinizing hormone in heifers.

Two experiments were designed to investigate the endocrine mechanisms by which progestin administration induces puberty in heifers. In experiment 1, prepubertal heifers were randomly assigned by age to receive either a single norgestomet implant for 10 days (NORG; n = 15) or to serve as unimplanted controls (CONT; n = 14). Serial blood samples were obtained on Days -0.5, 8.5, and 10.5 (Day 1 = day of implant insertion) and were analyzed for concentrations of LH. On days 9 and 11, 4 heifers in each treatment were slaughtered, and the reproductive tracts were obtained. Weekly progesterone analyses were performed to estimate the day of puberty in heifers not slaughtered. Puberty was induced in 6 of 7 heifers in the NORG treatment, resulting in an earlier (p < 0.05) day of puberty in the NORG than in the CONT treatment. The frequency of LH pulses was higher (p < 0.05) on Days 8.5 and 10.5 in the NORG as compared to the CONT treatment. Although no difference (p > 0.10) was observed between treatments in follicular development, uterine weight was greater (p < 0.05) in NORG than in CONT heifers on Day 11. In experiment 2, prepubertal heifers (n = 47) were administered either 1 (1NORG; n = 16), 3(3NORG; n = 16), or 0 (CONT; n = 15) norgestomet implants for 10 days, and serial blood samples were obtained as described for experiment 1. Transrectal ultrasonography was used to determine the diameter of the largest follicle on Day 9. Plasma samples were obtained after each serial sample collection period and were analyzed for estradiol concentrations. Puberty was induced in 75% (12 of 16) and 81% (13 of 16) of heifers in the 1NORG treatments, respectively. Four heifers in the 1NORG treatment, from which serial blood samples were collected, ovulated before removal of the progestin implant, and the LH data for this treatment were deleted. In the 3NORG treatment, LH pulse frequency was suppressed (p < 0.05) on Day 8.5, but was greater (p < 0.05) 12 h after removal of the progestin (Day 10.5) than in the CONT treatment. We conclude that progestin administration hastens puberty by increasing LH secretion after progestin withdrawal and propose that progestin administration induces puberty by accelerating the peripubertal decrease of estradiol negative feedback on LH secretion.

Animals↗

PubMed1995

Overexpression of bcl-2 protects prostate cancer cells from apoptosis in vitro and confers resistance to androgen depletion in vivo.

Normal (nonneoplastic) human prostatic secretory epithelial cells do not express the bcl-2 protein. However, a recent immunohistochemical survey of neoplastic human prostate tissues showed that a fraction of primary untreated prostate adenocarcinoma cells expressed this apoptosis-suppressing oncoprotein at significant levels (Colombel et al., Am. J. Pathol., 143: 390-400, 1993). Additionally, a number of hormone-refractory prostatic adenocarcinomas obtained from hormonally-treated patients (subsequent to surgical or drug castration therapy) were examined and were found to be uniform in their elevated expression of bcl-2 oncoprotein. The results of this preliminary survey imply that bcl-2 expression distinguishes a subgroup of primary human prostate cancers and that the expression of this protein might be a factor enabling prostate cancer cells to survive in an androgen-deprived environment. The current study was undertaken to determine the degree to which overexpression of bcl-2 can protect human prostate cancer cells from apoptotic stimuli in vitro and in vivo. Human prostate cancer cells (LNCaP) were transfected with a neomycin-selectable eucaryotic expression vector containing cDNA encoding human bcl-2. Transfected clonal variants that express bcl-2 protein (LNCaP/bcl-2) were unaltered with regard to their basal growth rate in 10% serum-containing medium, or with regard to their expression of the differentiated human prostate cell gene products prostate-specific antigen or androgen receptor protein. The bcl-2-transfected clones were altered, however, with regard to their growth rate in charcoal-stripped serum lacking dihydrotestosterone. Additionally, in contrast to the parental or control-transfected cell lines, LNCaP/bcl-2 cells were highly resistant to a variety of apoptotic stimuli in vitro including serum starvation and 10 nM phorbol ester (phorbol 12-myristate 13-acetate) supplementation of the medium. Lastly, the overexpression of bcl-2 by these prostate cancer cells altered their tumorigenic potential in a nude mouse assay. s.c. injections of 10(6) LNCaP/bcl-2 cells into male nude mice resulted in earlier and larger tumor formation compared to an equivalent injection of parental or control-transfected LNCaP cells. When these variant cell lines were injected into castrated male nude mice, only the LNCaP/bcl-2-transformed cells gave rise to tumors. Moreover, LNCaP/bcl-2 tumors grown in intact male nude mice were refractory to the growth-inhibiting effects of castration demonstrated by parental LNCaP cells. Data obtained in this study demonstrate that the bcl-2 oncoprotein can protect prostate cancer cells from apoptotic stimuli in vitro and suggest that such protection correlates with the ability to form hormone-refractory prostate tumors in vivo.

Androgen Antagonists↗

PubMed1995

Effects of limit feeding a high-grain diet on puberty and reproductive performance of ewes.

In Exp. 1, 39 ewe lambs (initial BW 29 +/- .1 kg) were used in a 2 x 2 factorial arrangement with two breeds (Polypay and Targhee) and two diets (high-forage and high-grain) to examine the effects of restricting intake of a high-grain diet on onset of puberty and lamb performance. The experimental diets were fed from 13 to 40 wk of age. The high-forage diet was composed of 80% alfalfa cubes and 20% supplement. The high-grain diet was 90% concentrate and 10% alfalfa cubes. Feed intake of the diets was controlled so that lambs fed the high-grain diet had the same growth rate as those fed the high-forage diet. Plasma insulin concentration was higher (P < .001) for lambs fed the high-grain diet than for those fed the high-forage diet. However, no diet effect on age at puberty was observed. In Exp. 2, 88 mature Targhee ewes were used to determine the effects of limiting intake of a high-grain diet on pregnancy and number of lambs born per ewe. Intake of the high-grain diet was restricted to achieve similar ME intake between diets. There were no diet effects on number of lambs born per ewe or subsequent lamb performance. Ewes fed the high-forage diet had a lower (P < .001) weight gain than those fed the high-grain diet. It was concluded that restricting intake of high-grain diets is an effective alternative to feeding high-forage diets for meeting the nutrient requirements of ewe lambs and gestating ewes; however, no improvements on reproductive performance were observed.

Animal Feed↗

PubMed1995

Hepatocyte growth factor and its receptor (c-MET) in prostatic carcinoma.

Hepatocyte growth factor (scatter factor) and its receptor, the c-met proto-oncogene product (c-MET), have been implicated in embryogenesis, tissue reorganization, and tumor progression. Little is known, however, of the expression and functional significance of these molecules in prostatic cells and tissue. In this investigation, we assessed the expression of hepatocyte growth factor (HGF) and c-MET in prostatic tissues and cell lines and also determined the effect of purified recombinant HGF on cell proliferation and scattering of prostatic carcinoma cell lines. HGF was expressed by human prostatic stromal myofibroblasts in primary culture but not by three human prostatic carcinoma cell lines (LNCaP, DU 145, and PC-3) as assessed by Northern blot analysis. HGF was also detected by reverse transcriptase-polymerase chain reaction in both benign and malignant tissues from radical prostatectomy specimens. c-MET transcripts were identified by Northern blot in two androgen-insensitive human prostatic carcinoma cell lines (DU 145 and PC-3) but not the androgen-sensitive LNCaP cell line. Additional evidence of linkage of androgen responsiveness and c-MET was provided by experiments in which androgen deprivation of normal rat prostates via castration produced a marked up-regulation of c-MET expression as determined by Northern blot and immunohistochemistry. c-MET protein was detected by immunohistochemical analysis in a substantial percentage (58 of 128 or 45%) of prostatic carcinomas and was found more often in metastatic growths of human prostatic carcinoma (15 of 20 patients) compared with primary tumors (43 of 108 patients; P < 0.005). Moreover, in Dunning R-3327 rat prostatic carcinoma cell lines, c-MET expression was highest in the androgen-insensitive subline with the highest metastatic capacity. Purified recombinant human HGF induced dose-dependent cellular proliferation and scattering in the DU 145 carcinoma cell line. These data indicate that HGF may function in the prostate gland as a paracrine growth factor, with synthesis by stromal cells and with biological target cells being the epithelial cells. Expression of the HGF receptor, c-MET, is up-regulated by androgen deprivation and c-MET appears to be preferentially expressed on androgen-insensitive, metastatic cells, suggesting a possible linkage of c-MET expression with prostatic carcinoma progression.

Animals↗

PubMed1994

The nuclear localization signal of NGFI-A is located within the zinc finger DNA binding domain.

NGFI-A is an immediate-early gene that encodes a transcription factor whose DNA binding domain is composed of three C2H2 zinc fingers. To identify its nuclear localization signal (NLS), wild type NGFI-A and various mutants were transfected into COS cells and their cellular location assayed by indirect immunofluorescence. Although wild type NGFI-A was located exclusively within the nucleus, deletions lacking the highly basic zinc finger region were not efficiently translocated to the nucleus. However, DNA binding per se is not required for nuclear localization, as an NGFI-A mutant (A Y339G), which does not bind DNA, is still faithfully directed to the nucleus. To determine the minimal region(s) of NGFI-A sufficient to direct nuclear localization, the cellular location of various NGFI-A/beta-galactosidase fusion proteins was examined. Fusion proteins containing all three zinc fingers were found in the nucleus, but those containing only two zinc fingers were predominantly cytoplasmic. When the zinc finger structure was altered by mutating a zinc-chelating cysteine residue in any one of the three zinc fingers, the resulting domain was no longer capable of directing beta-galactosidase to the nucleus. Furthermore, the mutation of an arginine residue in the third zinc finger of NGFI-A, a position which is occupied by a leucine residue in most C2H2 zinc fingers, abolished nuclear localization, but had no effect on DNA binding. These studies suggest that NGFI-A contains a novel NLS which is dependent on the overall structure of the DNA binding domain and not solely upon its highly basic nature.

Amino Acid Sequence↗

PubMed1994

Acute progesterone administration regresses persistent dominant follicles and improves fertility of cattle in which estrus was synchronized with melengestrol acetate.

Experiments were conducted to determine whether acute progesterone administration would regress persistent follicles and improve fertility in heifers and postpartum cows fed melengestrol acetate (MGA). In Exp. 1, heifers (n = 13) were fed MGA for 11 d (1st d of MGA = d 1). Prostaglandin F2 alpha (PGF2 alpha) was administered to heifers on d 2 to regress the corpus luteum (CL). On d 9, heifers were randomly assigned to receive an injection of either 200 mg of progesterone (PROG) or no hormone (vehicle; VEH). Neither growth of the persistent follicle nor plasma estradiol concentrations were altered by administration of VEH, and the persistent follicle ovulated after cessation of MGA feeding. Administration of PROG regressed the persistent follicle, reduced (P < .05) systemic estradiol concentrations, and resulted in ovulation of a newly recruited follicle. In Exp. 2, heifers were fed MGA for 14 d and were administered either PROG (n = 30) or VEH (n = 31) on d 12. When the CL was absent on d 12, synchronization conception rate (SCR) and pregnancy rate (PR) were greater (P < .05) for heifers administered PROG than for those administered VEH. Neither SCR nor PR were different among treatments when the CL was present on d 12. In Exp. 3, cows (n = 49) were fed MGA for 14 d and were administered either PROG or VEH on d 12. In cows lacking a CL (n = 32), administration of PROG increased (P < .05) SCR, but not PR. We conclude that acute PROG administration induces turnover of persistent follicles and may increase fertility when estrus is synchronized with MGA.

Animals↗

PubMed1994

Phorbol ester-induced apoptosis is accompanied by NGFI-A and c-fos activation in androgen-sensitive prostate cancer cells.

We have previously demonstrated the induction of the early response transcription factor NGFI-A in castration-induced regression of the rat ventral prostate. We have developed an in vitro model to investigate the role of kinase signal transduction and early transcriptional regulation in apoptosis of androgen-sensitive prostate cells. Cell death was induced in the androgen-sensitive human prostate line, LNCaP, by addition of the protein kinase activator, 12-tetradecanoylphorbol 13-acetate (TPA). TPA-induced death of LNCaP cells was asynchronous and exhibited morphological and ultrastructural features indicative of apoptosis. Specifically, cytoplasmic contraction, condensation of nuclear chromatin, and formation of membrane-bound "apoptotic bodies" were observed. Additionally, the characteristic endonuclease-generated DNA ladder, commonly associated with apoptosis, was observed in TPA-treated LNCaP cultures. TPA-induced LNCaP apoptosis was preceded by rapid yet transient induction of the early response transcription factors NGFI-A and c-fos. In dose-response experiments, NGFI-A and c-fos gene activation parallels the induction of death in LNCaP cells. TPA-induced expression of NGFI-A and c-fos and death of LNCaP cultures were blocked by pretreatment with staurosporine, a potent inhibitor of several protein kinases. Based on these studies, we suggest that activation of a TPA-inducible kinase(s) mediates apoptosis of androgen-sensitive prostate cells by means of an intracellular pathway that may involve the transient activation of the early response transcription factors NGFI-A and c-fos.

Alkaloids↗

PubMed1993

Prostatic nerve growth factor inducible A gene binds a novel element in the retinoblastoma gene promoter.

We are investigating the role of the early response transcription factor, nerve growth factor inducible A gene (NGFI-A), as a modulator of retinoblastoma (RB) gene transcription in prostate cells. Examination of the RB promoter reveals a novel element GCGGGGGAG located at nucleotides 152-144 upstream of the methionine initiation codon. This sequence shares strong homology with the consensus NGFI-A binding element GCGGGGGCG varying by a single nucleotide. In DNA binding assays, an NGFI-A fusion protein and the native protein product of the NGFI-A gene purified from prostate cancer cells bound specifically to an oligonucleotide containing the RB promoter element. Gene expression studies in rat ventral prostate demonstrated a 1.9-fold increase in RB mRNA following castration that parallels a 2.7-fold induction of NGFI-A mRNA. In summary, the in vitro DNA binding data and the transient coregulation of rat NGFI-A and RB following castration suggests that the RB gene may be transcriptionally regulated by NGFI-A in prostate cells.

Animals↗

PubMed1993

Cell-cycle control of a large-conductance K+ channel in mouse early embryos.

There have been few investigations into the role of ion channels in mammalian early embryonic development, despite studies showing that changes in ion channel activity accompany the early embryonic development of non-mammalian species and the proliferation of mammalian cells. Here we report that a large-conductance, voltage-activated K+ channel is active in unfertilized mouse oocytes but is rarely observed in later embryos. The channel activity is linked to the cell cycle, being active throughout M and G1 phases, and switching off during the G1-to-S transition. These changes in channel activity are accompanied by corresponding shifts in membrane potential. Inactivation of the channel during S/G2 can be prevented by exposing the oocytes to dibutyryl cyclic AMP or forskolin, an activator of adenylyl cyclase. Inhibition of protein synthesis with puromycin did not prevent inactivation of the channel at the end of G1 or its subsequent reactivation at the end of G2, indicating that the channel activity is not regulated by mitosis-promoting factor or cyclins.

Animals↗