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Biomedical subjects

R E Garfield

Publications and source records attributed to R E Garfield.

At least 127 records · Page 7Linked to original sources
PubMed1991

Effect of gap junction number and permeability on intercellular coupling in rat myometrium.

Gap junctions (GJ) increase between myometrial cells immediately before labor. To provide evidence of their role in cell-to-cell coupling, we evaluated input resistance (Ro) and intercellular spread of Lucifer yellow (LY) in intact preparations of rat longitudinal myometrium of preterm, term, and antiprogesterone-treated preterm delivering animals. LY injected into cells from either term or preterm delivering rats (many GJ) spread rapidly to neighboring cells by 60 s, but in preparations from nondelivering controls required 4-6 min to become detectable in adjacent cells. Ro of cells in preterm nondelivering preparations was 24.1 +/- 0.8 (SE) M omega, but dropped to 12.0 +/- 0.4 M omega (P less than 0.05) at delivery, similar to preterm delivering tissues at 13.8 +/- 0.6 M omega. The putative GJ uncoupling agent octanol reversibly increased Ro of term- and preterm-delivering tissues fourfold (P less than 0.01) within 60 s, and Ro of preterm-nondelivering tissue was further increased so that Ro values were similar among the three classes. These increased Ro values are interpreted as decreased coupling. Both K+ depolarization and oxytocin (10(-8) to 10(-6) M) increased Ro of delivering tissues (P less than 0.05), suggesting that high levels of contractile agonists may lead to reduced cell-to-cell coupling. Therefore, myometrial coupling can be modulated over seconds via GJ permeability as well as over hours by GJ number.

Animals↗
PubMed1990

Intrauterine occupancy controls expression of the parathyroid hormone-related peptide gene in preterm rat myometrium.

The parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHRP) genes are members of a gene family. Whereas PTH is a classical peptide hormone, mounting evidence suggests that the PTHRP may have predominately local actions. We report here that the PTHRP gene is expressed in rat myometrium, with a major peak in PTHRP mRNA expression occurring in the 48 hr immediately preceding parturition. A similar peak in peptide content was found in tissue extracts by biological and immunological assays, but the PTHRP could not be detected in the peripheral circulation or in uterine vein plasma during late gestation. By in situ hybridization histochemistry, PTHRP mRNA was demonstrated in both the longitudinal and circular layers of smooth muscle but was absent in the endometrium. The rise in myometrial PTHRP mRNA in late gestation was dependent upon intrauterine occupancy; it was greatly reduced or absent in nongravid uterine horns. These findings indicate that the expression of the PTHRP gene in preterm myometrium is under the control of a local stimulus and suggest that the PTHRP may play a paracrine or autocrine role in the uterus during the antepartum period, possibly involving myometrial contractility.

Animals↗
PubMed1990

Functional innervation of the myometrium of Myotis lucifugus, the little brown bat.

Myometria of pregnant and nonpregnant Myotis lucifugus were studied in vitro by using electrical field stimulation as well as autonomic agonists and antagonists to determine whether functional responses corresponded with structural evidence showing abundant adrenergic and sparse cholinergic innervation, which uniquely does not disappear during pregnancy. Field stimulation (70 V, 0.6 ms, 5.0-s pulse train, 2.5 - 60 Hz) of myometria from nonpregnant (hibernating) bats produced graded responses consisting of an initial alpha-adrenergic contraction and a subsequent beta-adrenergic relaxation phase. Responses were sensitive to both the nerve poison tetrodotoxin and the adrenergic antagonist guanethidine, demonstrating that they resulted from stimulation of intrinsic adrenergic nerves. Field stimulation responses were unaffected by atropine indicating that there was no functional cholinergic innervation, even though carbachol-induced contraction showed that muscarinic receptors were present. In contrast, functional innervation of cervical tissue was cholinergic and nonadrenergic-non-cholinergic, but not adrenergic. At the beginning of active gestation, some myometrial preparations exhibited little of no response to field stimulation. However, as uterine size increased, the biphasic response to field stimulation was enhanced, particularly the inhibitory (beta-adrenergic) phase. Moreover, the contractile phases, though reduced, was not abolished by alpha-adrenergic antagonists. The residual contractile response was also tetrodotoxin-resistant, suggesting that the myometrium was sensitive to direct electrical stimulation. Near the end of pregnancy, myometrial tissue became nonresponsive to both field stimulation and autonomic agonists, suggesting an absence of available receptor sites on muscle cells.

Animals↗
PubMed1990

Cell-to-cell coupling in the myometrium: Emil Bozler's prediction.

In this paper we have attempted to briefly review the studies which explain how the myometrium behaves as a functional syncytium. We believe that the gap junctions adequately explain the basis for cell-to-cell coupling as predicted by Emil Bozler fifty years ago.

Animals↗
PubMed1989

A functional and structural study of the innervation of the human uterus.

We characterized the innervation of human myometrial tissues by electrical field stimulation and electron microscopy. Nerve-specific parameters (pulse duration 0.6 msec) were used for electrical field stimulation to selectively activate intrinsic nerves. In specimens from nonpregnant, nonparous women (n = 6), tetrodotoxin (10(-6) mol/L) significantly reduced the response to electrical field stimulation by 70%. Contractions to electrical field stimulation were also inhibited to 60% by atropine (10(-5) mol/L) as well as by guanethidine (10(-5) mol/L) and phentolamine (10(-5) mol/L). Propranolol (10(-5) mol/L) had no detectable effect. We obtained similar results from about 50% of the specimens from nonpregnant, parous women (n = 15). The contractile responses of specimens from the term pregnant uterus (n = 13) to electrical field stimulation were not influenced by tetrodotoxin. Ultrastructurally we found nerve profiles in close proximity to muscle cells. About 30% of nerve varicosities in tissues from nonpregnant, nonparous patients could be classified as adrenergic (small, dense-cored vesicles), 53% as cholinergic (small, agranular vesicles), and about 17% as indeterminant (sometimes large, dense-cored vesicles). However, nerve varicosities were rarely observed in term pregnant specimens. These results indicate the presence of tetrodotoxin-sensitive, excitatory innervation of human myometrium consisting of alpha-adrenergic and cholinergic components. Furthermore, denervation may be nearly complete at term and recovery of innervation occurs at a considerable length of time after delivery.

Adult↗
PubMed1989

Increased myometrial responsiveness to oxytocin during term and preterm labor.

The contractile effects of oxytocin on the longitudinal and circular muscle of rat uteri from animals undergoing term and preterm labor were studied in vitro to define the action of the hormone. The maximal tension and slopes of oxytocin dose contractile-response curves, but not the pD2 values (-log ED50), were higher in both muscle layers at term delivery and also during preterm delivery induced with antiprogesterones compared with responses from tissues of nondelivering animals. These data indicate that the myometrium is hyperresponsive and hyperreactive to oxytocin during term and preterm birth but not hypersensitive. The increase in oxytocin responsiveness of the myometrium may be fundamental to the initiation of labor and regulated by the increases in oxytocin receptors and postreceptor events, including gap junctions.

Animals↗
PubMed1989

Mechanical responses of the rat uterus, cervix, and bladder to stimulation of hypogastric and pelvic nerves in vivo.

Mechanical activities of the uterus, cervix, and bladder were recorded in vivo in anesthetized rats during electrical stimulation of either the hypogastric or pelvic nerve. Ovariectomized controls and hormone-treated groups were used as well as pregnant and postpartum rats. Stimulation of either hypogastric or pelvic nerve produced voltage- and frequency-dependent contractions of the three organs with no evidence of apparent inhibition. All evoked responses were completely abolished by tetrodotoxin, suggesting that these nerves are common pathways of innervation to the three organs. Atropine abolished uterine and cervical responses to both hypogastric and pelvic nerve stimulation, whereas bladder responses were only partly reduced. Hexamethonium almost totally blocked the evoked responses of the uterus and cervix. Phentolamine partly blocked uterine and cervical responses, and propranolol or physostigmine enhanced uterine and cervical responses to both hypogastric and pelvic nerve stimulation. These results suggest that motor innervation to the rat uterus and cervix is predominantly postganglionic cholinergic, with some alpha- and beta-adrenergic components, and that the bladder is innervated by mainly cholinergic and also noncholinergic nerves. Estrogen and estrogen-plus-progesterone pretreatment significantly increased the responses of uterus and cervix but not bladder. Uterine and cervical responses to either hypogastric or pelvic nerve stimulation were markedly reduced late in pregnancy and reappeared within 7 days after delivery.

Animals↗
PubMed1989

Studies of the innervation of rabbit myometrium and cervix.

We characterized the innervation of isolated circular and longitudinal-oriented muscle strips from the nulliparous rabbit uterus and cervix by field stimulation (FS). FS with increasing frequency (2.5-50 pps) and voltage (2.5-70 V) caused graded increases in isometric contraction with no relaxation or inhibition of spontaneous activity. Tetrodotoxin (TTX, 3.1 X 10(-6) M) significantly reduced the FS response by 75% in all strips at higher stimulus frequencies. Contractile responses to FS were also significantly inhibited by atropine (3.5 X 10(-6) M) in circular uterus and in longitudinal cervix. Guanethidine (5 X 10(-6) M) reduced the response in all strips, as did phentolamine (3.6 X 10(-6) M) in longitudinal uterus and circular cervix. Propranolol (3.9 X 10(-6) M) did not significantly change the response in longitudinal uterus or circular cervix. In longitudinal uterus, combined guanethidine and atropine produced significant inhibition, but not statistically different from either drug alone. Similar results were seen in circular uterus. Electron microscopy and glyoxylic acid histofluorescence indicate that both blood vessels and smooth muscle in rabbit uterus are supplied with adrenergic nerves. The results suggest the presence of TTX-sensitive adrenergic and cholinergic excitatory innervation of rabbit uterus and cervix.

Adrenergic Fibers↗
PubMed1989

Improved propagation in myometrium associated with gap junctions during parturition.

The hypothesis that gap junction (GJ) formation between myometrial cells at term improves electrical coupling was tested. We measured the spread of electrical excitation from six extracellular electrodes aligned on uterine strips in either the longitudinal (axial) or transverse (circumferential) direction. Spontaneous bursts propagated over the entire 15-mm recording distance in the axial direction at both preterm and parturition and showed some characteristics of a system of coupled relaxation oscillators. However, individual spikes within the bursts propagated further and with higher velocity at parturition than at preterm. In the circumferential direction, both bursts and individual spikes propagated further at parturition than before. Propagation in this axis at parturition appeared to require an intact circular muscle layer. Spikes evoked by electrical stimulation also propagated further and with higher velocity in both axes at parturition. Electron microscopy showed many GJs between uterine smooth muscle cells during parturition, but few and sometimes no GJs at preterm. Thus improved propagation was associated with increased GJ contact between myometrial cells, consistent with the hypothesis that gap junction formation at term improves electrical coupling.

Action Potentials↗
PubMed1989

Density-dependent hyperpolarization in cultured aortic smooth muscle cells.

The membrane potential (Em) of cultured aortic smooth muscle cells from Sprague-Dawley (SD), Wistar-Kyoto (WKY), and spontaneously hypertensive (SHR) rats was measured in proliferating primary cultures. Em of SD cells in high-density cultures was -51 to -58 mV, whereas that of low-density cultures (1-2 days) was -30 mV. This difference was due to a continuous process of hyperpolarization during proliferation in culture. Em of WKY and SHR hyperpolarized similarly, from -12 to -42 and -38 mV, respectively. Hyperpolarization of Em of SD, WKY, and SHR cells was related to cell density rather than time in culture. Em may be a sensitive and significant indicator of the changes in the differentiated state expressed by proliferating smooth muscle in vitro.

Animals↗
PubMed1988

Innervation and gap junction formation in the myometrium of pregnant little brown bats, Myotis lucifugus.

Pregnant Myotis lucifugus were captured in mist nets set outside a large maternity colony and, in most cases, were examined 12-15 hours later. Anterior and posterior halves of uteri were pinned to dental wax and either incubated in glyoxylic acid to produce adrenergic nerve fluorescence, or fixed in buffered glutaraldehyde for electron microscopy. Blood was collected for radioimmunoassay of plasma progesterone. We found no evidence of decreased nerve fluorescence even in late pregnancy when plasma progesterone levels were 10-20 times preovulatory values. Ultrastructural examination also showed no evidence of damage to, or destruction of, nerves in the myometrium. However, we did find gap junctions between myometrial muscle cells during the periparturient period in both normal and aborting bats. Gap junctions began to form several days before term, increased in number until parturition, then decreased dramatically within a day or two thereafter. The percentage of muscle cell plasma membrane involved in gap junction formation was closely correlated with plasma progesterone levels, although whether this is causal or coincidental is not certain. These data do not agree with the conclusions drawn from observations in other mammals that a disappearance of adrenergic nerves from the myometrium is associated with the initiation of parturition, or that gap junction formation is associated with changes in nerve function. They do, however, lend further support to the hypothesis that there is neurogenic control of myometrial contractility in M. lucifugus uteri at term.

Animals↗
PubMed1988

Implant prostheses for convertibility, stress control, esthetics, and hygiene.

A method of connecting "fixed partial denture" prostheses to osseointegrated implant fixtures has been described. The advantages of this system of restoration for partially and fully edentulous mouths are that it is more effective in addressing the problems of (1) stress-control on abutments, (2) a back-up system for abutment failures, (3) esthetics, and (4) control of bacterial plaques around abutments. To accomplish this procedure, the application of convertible periodontal prosthesis techniques with modifications to some existing implant systems is undertaken. The disadvantages of this method seem insignificant when one considers the complexities and risks involved with the present array of implant prosthesis alternatives. Some patients and dentists might consider the necessity of the prosthesis being detachable as one disadvantage. In reality, the prosthesis can be used as a fixed restoration until the patient has fully adapted to the new proprioception and appearance. A large percentage of patients feel uncomfortable with the word "removable" because it immediately creates a perception of unsightly metallic clasp display, palatal coverage, tongue interference, and negative body image. The use of the term "detachable" coupled with the doctor's offer to perform this task for the patient "whenever necessary" will usually relieve the patient's anxiety and allow the treatment to proceed. Once neuromuscular and esthetic adaptation have occurred and the patient has accepted the prosthesis, daily detaching and home-care hygiene by the patient will follow without incident. Esthetic improvement is obvious (Fig. 3).

Cementation↗
PubMed1988

Effects of calcium ionophore, A23187 and calmodulin inhibitors on intercellular communication in the rat myometrium.

We have studied the effect of a calcium ionophore, A23187, and the purported calmodulin inhibitors, calmidazolium and chlorpromazine, on direct intercellular communication between smooth muscle cells in the myometrium of delivering rats. The extent of cell-to-cell coupling was determined by exposing one portion of small strips of longitudinal myometrium to 2-[3H] deoxy-D-glucose (2-DG) and determining the distribution and apparent diffusion coefficient (Da) for this tracer after a 5-h period for diffusion. The distribution and Da for 2-DG were significantly (p less than 0.05) reduced by exposure to A23187 in Krebs-Ringer solution with 2.5 mM Ca++, partially reduced in Krebs solution with A23187 and low Ca++ (1-10 microM), but the drug had no effect when used with Ca++-free solutions with [ethylenebis (oxyethylene-nitrilo)] tetraacetic acid (EGTA). The calmodulin inhibitors blocked the effects of A23187 in a dose-dependent fashion, and at higher concentrations, the extent of 2-DG diffusion was not different from that in control tissues. Surprisingly, however, a dose-dependent reduction in coupling was also observed in tissues exposed to the calmodulin inhibitors alone. Structural studies failed to reveal any change in the area of gap junctions between the myometrial cells following the above treatments, suggesting that the reduced exchange of 2-DG resulted from a decrease in the permeability of gap junctions between the muscle fibers.

Animals↗
PubMed1988

Control of myometrial contractility: role and regulation of gap junctions.

In this review we have attempted to describe the morphology and electrophysiology of the myometrium and also the myogenic, neurogenic and hormonal systems which control its activity. The synthesis of myometrial gap junctions appears to play a significant role in the gradual evolution of uterine contractility during labour. We suggest that the development of myometrial gap junctions is physiologically regulated by the steroid hormones, their receptors and prostaglandins. Furthermore, the permeability or conductance of the gap junction channels as well as their degradation seems to be hormonally regulated (Figure 10.6). Thus, there are several sites for the control of the presence and function of the gap junctions and the management of uterine contractility.

Animals↗
PubMed1987

In vitro studies on the control of human myometrial gap junctions.

In this study human myometrial tissues were examined for the presence of gap junctions by quantitative electron microscopy before and after incubation in tissue culture media with and without indomethacin. The area of gap junctions was very low in tissues from pregnant women at term but not labor, before incubation. After 24 and 48 h incubation without any treatment, segments of some of the same tissues developed many gap junctions and other tissues contained few junctions. Prostaglandin E (PGE), prostaglandin F (PGF) and prostaglandin F metabolite (PGF metabolite) levels in the media at various times were measured by radioimmunoassay. The prostaglandins increased progressively during the incubation period. Treatment of tissues with indomethacin decreased prostaglandin levels in the media and increased the numbers of gap junctions in those control tissues that developed few junctions over the same incubation interval. We conclude that the capacity of human myometrial tissues to develop gap junctions in vitro may depend upon a maturational stage in preparation for labor. Furthermore, our results suggest that products of the cyclo-oxygenase or lipoxygenase pathways may control the presence of gap junctions in the human myometrium and that changes in synthesis in these patterns may occur as part of the maturational process.

Female↗
PubMed1987

Effects of the antiprogesterone RU 486 on preterm birth in the rat.

The effects of the antiprogesterone compound RU 486 on preterm delivery, myometrial gap junctions, and plasma levels of steroid hormones were investigated in pregnant rats. Injection of RU 486 on day 16 of gestation resulted in prolonged delivery beginning after 24 hours. The preterm birth initiated by RU 486 was preceded and accompanied by the extensive development of gap junctions between myometrial cells. R 5020, but neither progesterone nor dexamethasone, prevented preterm birth and the development of gap junctions. Estrogen and progesterone levels declined after RU 486 treatment but only after a major proportion of the fetuses were delivered. This study indicates that progesterone may normally inhibit uterine contractility by suppressing the genome responsible for the expression of gap junctions.

Animals↗
PubMed1987

The antiprogesterone steroid RU 486: a short pharmacological and clinical review, with emphasis on the interruption of pregnancy.

In this review we have briefly outlined the clinical applications and mechanism of action of the progesterone antagonist RU 486. RU 486 has been successfully used in a variety of conditions to regulate the reproductive cycle and to control fertility in women. We suggest that the mechanism by which RU 486 acts during the cycle and early pregnancy is probably by affecting mainly the endometrium. During late pregnancy, the compound has significant effects on the myometrium including the induction of gap junctions between myometrium cells, which is required for muscle contractility during labour. The use of RU 486 has helped to demonstrate that progesterone is required for maintenance of the late stages of pregnancy in women.

Abortion, Induced↗