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Biomedical subjects

M Mattioli

Publications and source records attributed to M Mattioli.

At least 73 records · Page 4Linked to original sources
PubMed1991

Luteinizing hormone inhibits potassium outward currents in swine granulosa cells by intracellular calcium mobilization.

Potassium currents of swine granulosa cells were studied using the patch clamp technique in the whole cell configuration. Granulosa cells stepped to positive potentials (+60 mV) from -40 mV holding potential exhibit a slowly activating, noninactivating outward potassium current. Tail current reverse potential (between -90 and -100 mV) and the current inhibition brought about by the replacement of KCl with CsCl in the pipette solution indicate that this current is carried by K ions. LH was found to significantly reduce the amplitude of this current. The effect was dose and time dependent. Similar inhibition (20-30% of the initial current) was reached with doses of 1-50 micrograms/ml, but in times proportionally shorter as the dose increased (50% inhibition was reached in 170-180 and 30-40 sec with 1 and 50 micrograms LH/ml, respectively). Much longer and variable times (3-10 min) were required with lower doses (0.2 microgram/ml). The effect of LH was independent of extracellular Ca, while preexposure of cells to TMB-8, an inhibitor of intracellular Ca mobilization, completely prevented the effect of LH. Outward currents after LH treatment could be completely restored by perfusing the cells with ionomycin in Ca-free medium to facilitate calcium efflux from the cells. The present studies indicate that LH modifies the bioelectrical properties of swine granulosa cells. This effect is mediated by an elevation of intracellular calcium, probably mobilized from intracellular stores. The induced changes in K conductance may play a specific role in the transduction mechanisms for LH.

Animals↗
PubMed1990

The effect of antifreeze glycopeptides on membrane potential changes at hypothermic temperatures.

The research on antifreeze glycopeptides (AFGPs) from Antarctic and Arctic fishes has focused primarily on their interaction with ice crystals. This study reports results of experiments in which pig oocytes, known to be sensitive to hypothermic temperatures, were exposed to 4 degrees C for various periods of time, in solutions of different molecular weight AFGPs from Antarctic nototheniid fishes. The membrane potential was measured across the oolemma following hypothermic exposure. The results show that a physiological combination of the different molecular weight AFGPs protects the structural integrity of the oolemma and inhibits ion leakage across the oolemma at hypothermic temperatures. The results also show that the hypothermic protection is nonlinearly dependent on concentration and that separately, the different molecular weight glycopeptides do not stop ion leakage even at very high concentration. The protection of membranes at hypothermic temperatures is a new property of AFGPs which was not known prior to our work.

Animals↗
PubMed1990

Maturation of pig oocytes: observations on membrane potential.

The membrane-potential changes of pig oocytes during maturation are described. Cumulus-enclosed oocytes have a resting potential of -41.81 +/- 0.60 mV; the removal of cumulus cells caused this potential to drop to -30.95 +/- 0.43 mV. Adding LH to the culture medium did not influence the potential of denuded oocytes but depolarized the potential of cumulus-enclosed oocytes to -32.90 +/- 0.43 mV. FSH did not affect the membrane potential of denuded or cumulus-enclosed oocytes, but significantly reduced the amplitude of the depolarization induced by LH. The effect of gonadotropins on cultured granulosa cells was also investigated. Plated granulosa cells have a resting potential of -45.21 +/- 0.72 mV, similar to that of cumulus-enclosed oocytes. As recorded in cumulus-enclosed oocytes, LH depolarized granulosa cell membrane potential (-30.33 +/- 0.69 mV) and FSH reduced this effect. To evaluate if oocyte maturation in vivo is accompanied by membrane-potential depolarization, follicular growth and oocyte maturation were induced in 6 prepubertal gilts by using an eCG-hCG treatment. Twenty hours after the beginning of oocyte maturation in vivo (induced by hCG), the membrane potential of the oocyte was depolarized to -28.84 +/- 1.01 mV, a value similar to that observed in vitro. These data indicate that both LH and FSH can influence the membrane potential of follicular somatic cells and, consequently, that of the oocyte. The electrical coupling between somatic cell and oocyte may represent a means by which the gonadotropin message is passed to the germinal cell by the somatic compartment.

Animals↗
PubMed1989

Developmental competence of pig oocytes matured and fertilized in vitro.

Pig follicles 3 to 6 mm in diameter were everted and matured for 44 h. The oocytes were then collected and exposed to capacitated boar sperm purified by centrifugation in a two step (65 and 70%) Percoll gradient. Of 110 ova fixed 14 h after in vitro fertilization, 78% were penetrated and 47% were monospermic. Next, 681 oocytes were cultured in vitro for 44 h after in vitro fertilization and the 266 embryos which had reached the two- to four-cell stage were transferred into the oviducts of 12 synchronized recipient gilts. Four days later, 211 embryos (79%) were recovered by uterine flushing. 40.7% of these were at the blastocyst stage, and 20% were at the morula stage. In a final experiment, four out of eight gilts which had received 40 to 50 two- to four-cell embryos, were diagnosed pregnant 30 and 37 d after in vitro fertilization. One sow farrowed nine live piglets and one stillborn, two pregnancies were in progress, while one sow returned to estrus 47 d after in vitro fertilization. These results demonstrate that pig oocytes matured and fertilized in vitro can develop to the blastocyst stage and establish a normal pregnancy resulting in the birth of live piglets.

Journal Article↗
PubMed1988

Effect of follicle somatic cells during pig oocyte maturation on egg penetrability and male pronucleus formation.

In order to investigate the effect of the somatic cells of the follicle on the preparation of the oocyte for fertilization three experiments were carried out. In the first, pig oocytes, cultured for 46 h in the presence of extroverted follicles (follicle oocytes) or surrounded by the cumulus (cumulus oocytes), were exposed to sperm in an in vitro fertilization system. Follicle oocytes showed a higher rate of fertilization than that recorded in cumulus oocytes (80% vs. 47%). In addition, significantly more sperm penetrated into the ooplasm of follicular oocytes (3.77/egg) than into that of cumulus oocytes (1.42/egg). To investigate the reason for the observed fertilization difference zona-free oocytes were studied in the second experiment. Significantly more spermatozoa were recorded in the ooplasm of follicle oocytes than in that of cumulus oocytes, thus suggesting that the effect of the follicle on fertilizability may partly depend on an action on the plasma membrane of the oocyte. A further effect of the follicular tissue was on cytoplasmic maturation: only follicular oocytes were capable of consistently promoting male pronucleus formation. In cumulus oocytes, sperm that entered the cytoplasm remained in a condensed form near the female pronucleus. In the third experiment cumulus oocytes and denuded oocytes were matured in medium that had been previously used to mature follicle oocytes. This conditioned medium was alone able to affect sperm penetration and male pronucleus formation in cumulus oocytes, but it did not exert any influence on denuded oocytes. This suggests that the observed effect of the follicle is mediated by soluble factors that, however, cannot influence the oocyte without some direct cell-oocyte contract.

Animals↗
PubMed1988

Follicular factors influence oocyte fertilizability by modulating the intercellular cooperation between cumulus cells and oocyte.

In order to investigate whether the follicular tissue influences cumulus-oocyte interaction and, consequently, the fertilizability of the egg, four experiments were carried out. In the first, cumulus-enclosed pig oocytes were cultured for 44 h in control medium (modified TCM-199) or in follicle-conditioned medium, and the intercellular coupling was studied by measuring 3H-uridine uptake. In control medium the intercellular cooperation started to decline immediately, and at 24-32 h the uncoupling was almost complete. By contrast, in follicle, conditioned medium, it remained at high levels until 24-32 h. In the second experiment protein synthesis patterns of oocytes were studied. Oocytes cultured in conditioned medium were characterized by a 45-kD protein band, while those maturing in control medium were identifiable by a marked 56-kD band. In the third experiment mature oocytes were fertilized in vitro. The percentage of penetrated egg was higher in oocytes matured in conditioned medium than in control medium. In addition, only oocytes matured in conditioned medium could consistently decondense spermatozoa and form male pronuclei. Metabolic cooperation, protein synthesis patterns, and fertilizability were also studied in oocytes matured in control medium supplemented with either 17 beta-estradiol or progesterone or testosterone or dihydrotestosterone or androstenedione or ether extract of conditioned medium. Only ether extract and progesterone stimulated cumulus oocyte interaction and sperm decondensation. In the last experiment oocytes denuded at different stage of their maturation in conditioned medium were fertilized in vitro. The longer the eggs were cultured with the cumulus, the higher was their penetrability. Moreover, only oocytes denuded after 40 h of culture could, once fertilized, promote the formation of male pronuclei. These data demonstrate that follicular secretions are fundamental for the maintenance in vitro of a functional intercellular coupling between cumulus and oocyte, which is necessary for the egg to become penetrable by spermatozoa and to acquire the conditions required for the formation of male pronuclei.

Animals↗
PubMed1986

Effect of 5alpha-androst-16-en-3-one on oxytocin release in oestrous sows.

The effect of an aerosol containing 5alpha-androst-16-en-3-one on oxytocin release in the sow was studied. The snouts of six estrous sows were sprayed with the aerosol for 2 sec. Blood samples were collected through indwelling vena cava canulas. The aerosol released oxytocin in all the sows tested. Maximum values, ranging from 24 to 101 pg oxytocin/ml plasma were recorded within 8 min of the treatment. A possible positive effect of the pheromonal stimulation on pigs' fertility is hypothesized.

Journal Article↗
PubMed1986

The response of the coronary collateral circulation to acute administration of nifedipine: an angiographic and ergometric study.

To evaluate the role of collaterals in patients with effort angina we retrospectively compared the coronary cineangiograms of 14 subjects ("responders") who improved their exercise tolerance after acute nifedipine therapy with 14 subjects ("non-responders") with the same symptomatology who did not respond to the same treatment. The status of collaterals was graded with a score from a minimum of 0 to a maximum of 5. The responders showed a greater score than the non-responders (3 +/- 1 vs. 1 +/- 1, P less than 0.001), whereas there was no difference in the number of stenosed vessels between the two groups (1.8 +/- 0.9 vs. 2 +/- 0.8). Thus, in patients with effort angina and critical coronary stenosis, the presence of an efficient coronary collateral circulation can favour the increase in coronary flow reserve after vasodilator therapy. Our results suggest that the grading of collaterals may add useful information to the simple classification of one-, two- or three-vessel coronary artery disease.

Adult↗
PubMed1986

Effect of naloxone on plasma concentrations of prolactin and LH in lactating sows.

Six lactating sows were injected through an indwelling vena cava cannula with naloxone (2.5 mg/kg body weight) on Day 15 post partum. Blood samples were collected through the cannulas at 10-min intervals for 8 h before and 10 h after naloxone administration. Plasma prolactin and LH concentrations were measured by radioimmunoassay. Naloxone caused a marked suppression of plasma prolactin concentrations lasting 4-6 h. LH concentrations were also affected by naloxone: LH rose to reach maximum values 20-50 min after naloxone treatment. Pretreatment values were recorded 200-300 min after the treatment. These results indicate that endogenous opioids are involved in causing the endocrine patterns occurring during lactation, i.e. high prolactin and low LH concentrations.

Animals↗
PubMed1985

Effect of PGF-2 alpha on progesterone production in swine luteal cells at different stages of the luteal phase.

Suspensions of luteal cells were prepared by enzymatic dispersion of pig corpora lutea obtained at specific times during the estrous cycle. Luteal cells from early corpora lutea produced more progesterone (4.73 +/- 0.84 nmol/10(6) cells, day 3) than those from late diestrus (0.73 +/- 0.04 nmol/10(6) cells, day 15); (P less than 0.05). Bovine LH enhanced progesterone production in a dose dependent manner particularly in cells from 9 to 15 day corpora lutea. Also PGF-2 alpha enhanced progesterone output in cells from mid-late corpora lutea. PGF-2 alpha did not exert any antigonadotropic effect since it further increased the progesterone production induced by LH. Luteal cells produced PGF-2 alpha with levels ranging between 1.6 and 2.7 pmol/10(6) cells throughout the whole luteal phase. The cellular content of cAMP was markedly increased by LH (556 +/- 60%) while it was not affected by PGF-2 alpha. Plasma membrane receptors for PGF-2 alpha were not detected in the analyzed tissue.

Animals↗
PubMed1984

Increased sodium-lithium countertransport in red cells of patients with Bartter's syndrome.

Erythrocyte sodium-lithium countertransport was evaluated in three adult patients with Bartter's syndrome, diagnosed on clinical and histopathological basis and on laboratory tests. As compared with age-matched control subjects, lithium effluxes to both sodium medium and to sodium-free medium were high in two patients and Na+-Li+ countertransport was significantly greater in all three patients. The relationship between increased Na+-Li+ countertransport and the proposed primary disorder of Bartter's syndrome (i.e., a primary tubular defect of chloride reabsorption) is not apparent. However, this finding favours the hypothesis of a generalized cellular membrane dysfunction of ion transport in Bartter's syndrome.

Adult↗
PubMed1984

Androstenedione and testosterone concentrations in plasma and milk of the cow throughout pregnancy.

Androstenedione concentrations in both plasma and milk were higher than those of testosterone throughout pregnancy. Testosterone concentration increased during pregnancy; the levels in milk were always lower than in plasma. Androstenedione concentrations increased in plasma and milk during pregnancy but the values were twice as high in milk as in plasma after Day 90. This change in androstenedione distribution suggests that the mammary gland could be active in the uptake of androstenedione from plasma to milk or that the mammary gland itself might synthesize this hormone.

Androstenedione↗
PubMed1983

Characteristics of receptors for prostaglandin F-2 alpha in bovine and equine corpora lutea.

Prostaglandin F-2 alpha (PGF-2 alpha) receptors of bovine and equine corpora lutea (C.L.) were studied. From both the equilibrium binding data and the dissociation kinetics behaviour, two affinity classes of receptors are evident in the mare, with apparent dissociation constants (Kd) of 1,5 x 10(-9) M and 3.5 x 10(-8) M. Bovine PGF-2 alpha receptors present a homogeneous population of binding sites with Kd = 1 x 10(-8) M. Both bovine and equine C.L. receptors bind PGF-2 alpha in a specific manner; only 13, 14-dihydro-PGF-2 alpha considerably cross-reacts with these receptors. Since in the mare uterine PGF-2 alpha reaches the ovary systemically with consequent extensive degradation, it is suggested that this metabolite could act as a luteolysin in this species. The dissociation kinetics indicate that the hormone-receptor (H-R) reaction evolves from a state of loose binding to one of tight binding as the time of incubation proceeds. This could be explained by internalization of the H-R complex.

Animals↗