Biomedical subjects

D A Douglas

Publications and source records attributed to D A Douglas.

10 recordsLinked to original sources

PubMed1998

Differentiation of the corpus luteum of the mink (Mustela vison): mitogenic and steroidogenic potential of luteal cells from embryonic diapause and postimplantation gestation.

The mink corpus luteum (CL) involutes after ovulation and remains dormant, synthesizing low amounts of progesterone until reactivated to terminate embryonic diapause. We examined the mitotic and steroid synthetic capacity of luteal cells from the diapause and postimplantation phases of mink gestation. Cells from diapause divided in vitro, reaching confluence in 7-8 days. Three phenotypes were distinguishable: a fusiform cell in whorls, a hypertrophied epithelioid cell, and a small epithelioid cell. The first and second cell types divided in vitro after confluence, evidenced by localization of proliferating cell nuclear antigen (PCNA) in their nuclei. The small epithelioid cells were present in cell nests and showed no PCNA activity. Cells derived from reactivated CL did not reach confluence and had no PCNA activity. Progesterone accumulation was enhanced in luteal cells from diapause by LH, FSH, and dibutyryl (Bu2)cAMP, and by LH and (Bu2)cAMP in cells from reactivated CL. In luteal cells from the diapause phase of gestation, LH and (Bu2)cAMP induced increases in mRNA coding for steroidogenic acute regulatory protein, while cytochrome P450 side-chain cleavage enzyme mRNA was increased by prolactin, LH and (Bu2)cAMP. Cellular concentrations of 3beta-hydroxysteroid dehydrogenase-delta5-4-isomerase mRNA were increased by prolactin and (Bu2)cAMP. Thus, luteinization in the mink CL does not engender exit from the cell cycle, as both fusiform and hypertrophied cells from diapause divide in vitro. Reactivation appears to represent terminal differentiation. LH is capable of stimulating steroidogenesis in vitro in luteal cells from diapause, and prolactin and LH appear to have both specific and overlapping stimulatory effects on the CL of this species.

Animals↗

PubMed1998

Luteotropic hormone receptors in the ovary of the mink (Mustela vison) during delayed implantation and early-postimplantation gestation.

The reproductive cycle of the mink displays rigid seasonality and obligate embryonic diapause. After ovulation, the corpus luteum (CL) involutes, and it secretes basal progesterone until activated prior to implantation. To study changes in the relative abundance of luteal prolactin and LH receptor mRNA through gestation, ovaries and serum were collected from pregnant female mink at 2-day intervals (n = 3 per date) through embryonic diapause and CL activation (March 19-31) and at 5-day intervals through implantation and early-postimplantation gestation (March 31-April 15). To determine the effect of endogenous prolactin, mink received Alzet osmotic minipumps releasing 2 mg/day 2-bromo-alpha-ergocryptine (bromocriptine) or saline on March 19. Ovaries and serum were taken from 3 animals every 2 days until March 31. Prolactin receptor mRNA in ovaries was low during CL activation but increased 3-fold through embryo implantation. Its abundance correlated with prolactin binding to ovarian membranes and with circulating prolactin. Bromocriptine suppressed endogenous prolactin levels and prevented the increase in prolactin receptor mRNA. There was a transient peak in LH receptor mRNA in the ovaries at March 19-23, which declined to basal levels by March 25 and remained constant through midgestation. Bromocriptine prevented the preimplantation peak in LH receptor mRNA and reduced its abundance below pretreatment levels. The results suggest that prolactin up-regulates its receptor and maintains the LH receptor in the mink CL. The pattern of LH receptor mRNA argues for a role for LH in CL reactivation and termination of embryonic diapause in mink.

Animals↗

PubMed1997

Computational sequence analysis of the tissue inhibitor of metalloproteinase family.

The tissue inhibitor of metalloproteinase (TIMP) family regulates extracellular matrix turnover and tissue remodeling by forming tight-binding inhibitory complexes with matrix metalloproteinases (MMPs). MMPs and TIMPs have been implicated in many normal and pathological processes, such as morphogenesis, development, angiogenesis, and cancer metastasis. This minireview provides information that would aid in classification of the TIMP family and in understanding the similarities and differences among TIMP members according to the physical data, primary structure, and homology values. Calculations of molecular weight, isoelectric point values, and molar extinction coefficients are reported. This study also compares sequence similarities and differences among the TIMP members through calculations of homology within their individual loop regions and the mature region of the molecule. Lastly, this report examines structure-function relationships of TIMPs. Thorough knowledge of TIMP primary and tertiary structure would facilitate the uncovering of the molecular mechanisms underlying metalloproteinase, inhibitory activities and biological functions of TIMPs.

Algorithms↗

PubMed1997

Luteal and placental characteristics of carnivore gestation: expression of genes for luteotrophic receptors and steroidogenic enzymes.

Experiments were carried out to investigate the abundance of mRNA for luteotrophic receptors and steroidogenic elements in the ovaries and corpora lutea of mink during the embryonic diapause, peri-implantation and postimplantation pregnancy. The second aim was to determine whether the mink placenta synthesized progesterone. Homologous cDNA probes for the mink LH and prolactin receptors were generated by the polymerase chain reaction. Heterologous cDNA probes for steroidogenic acute regulatory protein (StAR), cytochrome P450 side chain cleavage (P450scc) and 3 beta-hydroxysteroid dehydrogenase-delta 4-delta 5 isomerase (3 beta HSD) were also used. The abundance of mRNA encoding the prolactin receptor was low during the period of embryonic diapause and increased concurrent with circulating progesterone. The abundance of LH receptor message reached peak values during the peri-implantation period followed by maintenance of a steady-state after implantation. The abundance of StAR and P450scc messages appeared not to vary during gestation, while that for 3 beta HSD was correlated with changes in circulating progesterone. There was no evidence of 3 beta HSD activity or transcripts in the placenta. These results indicate that prolactin and LH are necessary for activation of the corpus luteum during the period of embryonic diapause, and for its maintenance during postimplantation gestation. The mink placenta does not synthesize progesterone.

3-Hydroxysteroid Dehydrogenases↗

PubMed1996

Computational sequence analysis of matrix metalloproteinases.

Matrix metalloproteinases (MMP) play a cardinal role in the breakdown of extracellular matrix involved in a variety of biological and pathological processes. Research on MMPs has classified and characterized these enzymes according to their matrix substrate specificity, gene and protein domain structure, and regulation of activity and expression. However, the discovery of new MMPs has introduced a need for a more comprehensive and systematic method of classification and quantitative comparison of known and newly discovered members. This study compiles a sequence alignment, constructs a dendrogram, and calculates physical data and homology percentage assignments in order to obtain further insight into MMP structure-function relationships. Thorough analysis of MMP primary sequence domains, physical data patterns, and statistical analysis of sequence homology yields higher resolution in the similarities and differences that group MMP members.

Algorithms↗

PubMed1996

Census variation staffing.

A Census Variation Staffing (CVS) model has been used successfully on all nursing units for 4 years. Historical data and nursing hours per patient day (NHPPD) are used to determine the staffing needs of each unit on a daily, shift-by-shift basis. CVS has been heralded as the single largest factor in the hospital's consistent profitability--averaging annual savings of $485,100.

Bed Occupancy↗

PubMed1995

Development of immortalized endometrial epithelial and stromal cell lines from the mink (Mustela vison) uterus and their effects on the survival in vitro of mink blastocysts in obligate diapause.

Mink endometrial cell lines were established by stable transfection of a plasmid vector encoding the SV40 large T antigen driven by the human beta-actin promoter. A second plasmid vector, pSV2neo, was employed for selection of transfected cells. Specificity and homogeneity of consequent cell lines were evaluated by immunocytochemistry employing antibodies against cytokeratin, desmin, and vimentin. Cytokeratin was found exclusively in epithelial cells, whereas vimentin appeared primarily in stromal cells. Neither cell line showed detectable desmin activity. These cell lines along with Buffalo rat liver (BRL) cells were employed in coculture with mink embryos in obligate diapause. Mink stromal and BRL cell lines were most effective in enhancing embryo survival in vitro. The percentages of cocultured embryos that survived for 72 h or more were 65% with epithelial cells, 75% with stromal cells, 68% with the combination of stromal and epithelial cells, and 93% with BRL cells. Only 23% of the embryos cultured without cells survived beyond 48 h. Embryo growth was also observed; some embryos in coculture showed trophoblastic outgrowth and adhesion to the cell surfaces. These results demonstrate that mink embryos in obligate delay can survive and develop in culture and that coculture with uterine or BRL cells increases the length and frequency of survival.

Alkaline Phosphatase↗

PubMed1994

Ovarian follicular development in mink (Mustela vison).

Ovarian follicular dynamics were studied during the breeding season, before and after ovulation in mink. Nulliparous female mink were stimulated to ovulate with an injection of 4 micrograms GnRH. Ovaries from three animals were collected on days 0, 2, 3, 4, 5, 6 and 7 after hormone treatment. A second dose of GnRH was administered on day 8 and ovaries were collected from three animals on day 9. Corpora lutea and follicles were identified in histological sections and follicles were classified by stage of development, healthy versus atretic, and by diameter. Preovulatory follicles (diameter 0.7-1.0 mm) were present in the ovaries of all animals on day 0 and these responded to GnRH treatment by ovulating. A synchronized wave of follicular development occurred following ovulation. Changes in follicle populations indicated that follicles are recruited from the small antral follicle class (0.2-0.4 mm) into the 0.4-0.6 mm class, with the first defined changes occurring between days 2 and 4. From the recruited group, a smaller cohort of follicles is selected to become the dominant follicles between days 4 and 6, and these acquire the ability to respond to a stimulus which induces ovulation at diameters of > 0.7 mm. The ovaries of unmated mink also contained substantial numbers of large, degenerating, luteinized, unruptured follicles. These degenerating, luteinized follicles are considered to represent the demise of large follicles that failed to receive an ovulatory stimulus.

Animals↗

PubMed1993

Induction of follicular development in silver foxes (Vulpes vulpes) with equine chorionic gonadotrophin (eCG) and antibodies against eCG.

Experiments were conducted that demonstrated that 1000 iu equine chorionic gonadotrophin (eCG) was effective in induction of follicular development and ovulation in silver foxes during anoestrus. This treatment resulted in large, unovulated follicles; thus, trials in which the effects of eCG were reduced or abrogated by antibodies against eCG were carried out. Passive immunization against eCG on days 3 and 4 after eCG treatment interfered with subsequent follicular development and prevented ovulation. Treatment with eCG antibodies on days 5 or 7 after eCG treatment did not prevent ovulation and neutralization beginning on day 5 appeared to provide for the best ovulatory yield. The results suggest that combinations of eCG and anti-eCG antibody may provide a useful means of inducing ovarian activity in anoestrous foxes.

Anestrus↗

PubMed1990

Interactions between melatonin and prolactin during gestation in mink (Mustela vison).

Immature female mink, 8 weeks of age in July, were treated with implants releasing melatonin. Mating, which induced ovulation, took place during the normal breeding season in the following March. Circulating prolactin and progesterone concentrations did not undergo the expected gestational increases, and no embryos implanted. A similar absence of gestational changes in prolactin and progesterone values ensued in primiparous mink treated with the melatonin implant 2-3 days after the second of 2 matings. Administration of exogenous sheep prolactin (0.5 mg/day) by minipump induced precocious elevation of progesterone concentrations in mated mink. Prolactin administration overcame the effects of melatonin, in that the corpora lutea were activated and embryos implanted, but exogenous prolactin resulted in degeneration of implanted embryos both in the presence and absence of chronic melatonin. The results suggest that melatonin has a single effect in alteration of gestation in mink; i.e. the prevention of prolactin secretion. Hyperprolactinaemia may inhibit embryo development in this species.

Animals↗