THE LEVATOR ANI MUSCLE OF THE RAT AS AN INTACT PREPARATION SUITABLE FOR IN VITRO INVESTIGATIONS

1966 ◽  
Vol 52 (2) ◽  
pp. 325-336 ◽  
Author(s):  
A. Arvill ◽  
K. Ahrén
Keyword(s):  
Levator Ani ◽  
Male Rats ◽  
Levator Ani Muscle ◽  
List Type ◽  
Tissue Water Content ◽  
Tissue Water ◽  
Incubation Periods ◽  
Total Tissue

ABSTRACT A method is described of dissecting out and incubating the levator ani muscle of immature male rats, keeping its normal connections to the rest of the perineal complex. In order to find out whether this isolated preparation remained intact with undamaged cell membranes, comparisons were made between this preparation of the levator ani muscle and a cut preparation of the same muscle, and also between the levator ani muscle and the intact and cut preparations of the diaphragm. The following determinations were made: Distribution of sucrose-14C in vivo and in vitro Distribution of inulin-14C in vitro Distribution of D-xylose-14C in vitro Total tissue water content Potassium concentration in the medium after different incubation periods. It is concluded that it is possible to dissect out the levator ani muscle with undamaged cells and that this preparation can be both suitable and useful for in vitro investigations.

EFFECTS OF GROWTH HORMONE ON THE METABOLISM OF THE ISOLATED LEVATOR ANI MUSCLE OF THE RAT

1967 ◽  
Vol 56 (3_Suppl) ◽  
pp. S15-S25 ◽  
Author(s):  
A. Arvill ◽  
Å. Hjalmarson
Keyword(s):  
Growth Hormone ◽  
Distribution Ratio ◽  
Accumulation Rate ◽  
Levator Ani ◽  
Male Rats ◽  
Normal Male ◽  
Levator Ani Muscle ◽  
Hypophysectomized Rats

ABSTRACT The effects of growth hormone (GH) were investigated in the levator ani muscle and the diaphragm from hypophysectomized and from normal male rats. GH was added to the incubation media and/or injected daily for four days to the rats before the in vitro experiments. GH, added in vitro, stimulated the rate of accumulation of D-xylose-14C in the two muscles from hypophysectomized but not from normal rats. GH, in vitro, increased the distribution ratio of α-aminoisobutyric acid-14C (AIB-14C) in the diaphragm from hypophysectomized rats to a much higher degree than in muscles from normal rats. In vivo injections of GH to hypophysectomized rats which were not stimulating in itself, made the diaphragm insensitive to GH added in vitro. In the levator ani muscle, however, a different effect was obtained on the AIB-14C distribution ratio. GH in vitro did not stimulate muscles from hypophysectomized rats more than those from normal rats. Furthermore, administration of GH to hypophysectomized rats, which not was stimulating in itself, gave an additional effect with GH in vitro in this muscle. The accumulation rate as well as the incorporation into protein of glycine-3H increased in both muscles from hypophysectomized rats after in vivo as well as in vitro administration of GH. The results are discussed in connection to our present knowledge of the different effects of GH.

Espace extracellulaire, perméabilité à l'inuline et accumulation de L-alanine dans l'intestin isolé de Grenouille

1973 ◽  
Vol 51 (1) ◽  
pp. 22-28
Author(s):  
Joël de la Noüe ◽  
André Gagnon
Keyword(s):  
Extracellular Space ◽  
Muscle Layer ◽  
Solute Concentration ◽  
Tissue Water ◽  
Intracellular Amino Acid ◽  
Inulin Space ◽  
Total Tissue ◽  
Frog Intestine

In order to calculate the intracellular concentration of accumulated L-alanine, the extracellular space (inulin-14C) of frog intestine was measured. To check the validity of the technique, frog liver and gastrocnemius were used too. By scraping proximal portions of intestine, the inulin space was found to be similar (around 20% of total tissue water) in both the muscle layer and the mucosa. The mucosal epithelium is an imperfect barrier to inulin while the serosa is very permeable. These results suggest that the interstitial solute concentration is best approximated by equating it to that of the serosal solution. The in vitro inulin space, compared to the in vivo one, increases with time, as does the cellular hydration. The data obtained from measurements of extracellular space and from L-alanine uptake show that the intracellular amino acid is in a free state.

Hemorrhagic shock impairs myocardial cell volume regulation and membrane integrity in dogs

1987 ◽  
Vol 252 (6) ◽  
pp. H1203-H1210
Author(s):  
J. W. Horton
Keyword(s):  
Hemorrhagic Shock ◽  
Cell Volume ◽  
Volume Regulation ◽  
Membrane Integrity ◽  
Cell Volume Regulation ◽  
Calcium Entry ◽  
Tissue Water ◽  
Total Tissue

An in vitro myocardial slice technique was used to quantitate alterations in cell volume regulation and membrane integrity after 2 h of hemorrhagic shock. After in vitro incubation in Krebs-Ringer-phosphate medium containing trace [14C]inulin, values (ml H2O/g dry wt) for control nonshocked myocardial slices were 4.03 +/- 0.11 (SE) for total water, 2.16 +/- 0.07 for inulin impermeable space, and 1.76 +/- 0.15 for inulin diffusible space. Shocked myocardial slices showed impaired response to cold incubation (0 degrees C, 60 min). After 2 h of in vivo shock, total tissue water, inulin diffusible space, and inulin impermeable space increased significantly (+19.2 +/- 2.4, +8.1 +/- 1.9, +34.4 +/- 6.1%, respectively) for subendocardium, whereas changes in subepicardium parameters were minimal. Shock-induced cellular swelling was accompanied by an increased total tissue sodium, but no change in tissue potassium. Calcium entry blockade in vivo (lidoflazine, 20 micrograms X kg-1 X min-1 during the last 60 min of shock) significantly reduced subendocardial total tissue water as compared with shock-untreated dogs. In addition, calcium entry blockade reduced shock-induced increases in inulin impermeable space and inulin diffusible space. In vitro myocardial slice studies confirm alterations in subendocardial membrane integrity after 2 h of in vivo hemorrhagic shock. Shock-induced abnormalities in myocardial cell volume regulation are reduced by calcium entry blockade in vivo.

COMPARISON BETWEEN THE BINDING OF 19-NORTESTOSTERONE, 5α-DIHYDROTESTOSTERONE AND TESTOSTERONE IN RAT PROSTATE AND BULBOCAVERNOSUS/LEVATOR ANI MUSCLE

1976 ◽  
Vol 70 (3) ◽  
pp. 379-387 ◽  
Author(s):  
M. KRIEG ◽  
M. DENNIS ◽  
K. D. VOIGT
Keyword(s):  
Specific Binding ◽  
Levator Ani ◽  
Levator Ani Muscle ◽  
Binding Affinities ◽  
Receptor Sites ◽  
Rat Prostate ◽  
Layer Chromatography ◽  
In Vitro Data

SUMMARY Specific binding of [3H]19-nortestosterone in the 100000 g cytosol of the rat bulbocavernosus/levator ani muscle (BCLA) and prostate was demonstrated by agargel electrophoresis at low temperature and compared qualitatively and quantitatively with the binding of tritiated testosterone and 5α-dihydrotestosterone (5α-DHT). Both tissues showed a greater binding affinity for 5α-DHT than for 19-nortestosterone, with testosterone binding the least well of the three. The relative binding affinities in the BCLA and prostate were: 19-nortestosterone: testosterone = 1·4, 19-nortestosterone: 5α-DHT = 0·7. The differences were statistically significant (P < 0·02). The concentrations of receptor sites for 5α-DHT were 171 ± 20 (s.d.) fmol/mg prostatic cytosol protein and 24 ± 4 (s.d.) fmol/mg BCLA cytosol protein. The in-vitro metabolism of the three steroids in both tissues was also investigated by thin-layer chromatography. After incubating for 2 h at 0 °C the prostate was shown to reduce 26% of the 5α-DHT to androstanediols whilst the BCLA showed a 5% conversion. Testosterone was converted by the prostate to 5α-DHT (10%) and the androstanediols (6%) whilst the BCLA showed little activity in this respect. Comparing these in-vitro data with in-vivo findings from the literature, in both organs there is a positive correlation of the extent of binding in vitro to the stimulation of growth in vivo, bearing in mind that testosterone is metabolized to 5α-DHT in the prostate whilst in the BCLA, 5α-reductase is essentially absent.

DIFFERENTIAL EFFECTS OF CASTRATION AND DENERVATION ON PROTEIN SYNTHESIS IN THE LEVATOR ANI MUSCLE OF THE RAT

1972 ◽  
Vol 54 (1) ◽  
pp. 3-NP ◽  
Author(s):  
M. BUREŠOVÁ ◽  
E. GUTMANN ◽  
V. HANZLÍKOVÁ
Keyword(s):  
Protein Synthesis ◽  
Levator Ani ◽  
Mature Male ◽  
Male Rats ◽  
Levator Ani Muscle ◽  
Female Rat ◽  
Weight Changes ◽  
Immature Male ◽  
Testosterone Administration

SUMMARY The effects of testosterone on weight changes, incorporation of labelled precursors into RNA and proteins, and on ultrastructure of the levator ani muscle of male rats in which castration or denervation was performed, were studied. No increase of weight occurred in the denervated levator ani muscle after castration. There was no increase in the incorporation of labelled precursors into RNA and proteins in vitro in mature rats, if the muscle was denervated. However, incorporation still increased when denervation was performed in sexually immature animals. The levator ani muscle, which normally undergoes postnatal involution in the female rat, could be maintained even after postnatal denervation if testosterone was administered at birth, the hormone acting as a nerve-independent trophic agent. The increase of protein synthesis in the levator ani of mature, male, castrated animals, induced by testosterone administration, was abolished after denervation of the muscle. The activating influence of testosterone upon protein synthesis was, however, still present in immature male animals.

MAMMOGENIC EFFECTS OF METHANDROSTENOLONE IN CASTRATED RATS

1963 ◽  
Vol 42 (4) ◽  
pp. 601-614 ◽  
Author(s):  
K. Ahrén ◽  
A. Arvill ◽  
Ä. Hjalmarson
Keyword(s):  
Levator Ani ◽  
Mammary Glands ◽  
Seminal Vesicles ◽  
Female Rats ◽  
Male Rats ◽  
Weight Increase ◽  
Levator Ani Muscle ◽  
List Type ◽  
Alveolar Development ◽  
Dose Levels

ABSTRACT Methandrostenolone (17β-hydroxy-17α-methyl-androsta-1,4-dien-3-one) was injected in 4 dose-levels (0.05, 0.5, 2.5 and 5.0 mg daily for 28 days) into castrated male rats, and the response of the mammary glands, the seminal vesicles and the levator ani muscle studied. One dose-level (2.5 mg daily for 28 days) was injected into castrated female rats, and the response of the mammary glands, the vagina and the uterus studied. The main results were as follows: The 0.05 mg dose did not stimulate the seminal vesicles but produced a slight weight increase of the levator ani muscle. The 0.5 mg dose had only a minimal effect on the seminal vesicles but had a much more pronounced effect on the levator ani muscle. The 2 higher doses, however, markedly stimulated both the seminal vesicles and the levator ani muscle. In the mammary glands methandrostenolone produced not only lobule-alveolar development, as do most other androgens, but in addition induced growth and development of the mammary duct system, as found with oestrogenic compounds. The lobule-alveolar development, found after treatment with the various dose-levels of methandrostenolone, quantitatively, more closely followed the growth of the levator ani muscle than the development of the seminal vesicles. In the castrated female rats methandrostenolone stimulated vaginal opening, brought about slight cornification of the vaginal epithelium and caused a marked weight increase of the uterus. These effects cannot be explained solely on the basis of the androgenic activity of this compound, but seem to indicate that methandrostenolone has an oestrogenic activity when injected into castrated rats.

EFFECT OF TESTOSTERONE ON PROTEIN SYNTHESIS AND CONTRACTILITY OF THE LEVATOR ANI MUSCLE OF THE RAT

1971 ◽  
Vol 50 (4) ◽  
pp. 643-651 ◽  
Author(s):  
M. BUREŠOVÁ ◽  
E. GUTMANN
Keyword(s):  
Protein Synthesis ◽  
Latency Period ◽  
Previous Treatment ◽  
Levator Ani ◽  
Diaphragm Muscle ◽  
Levator Ani Muscle ◽  
Contraction Time

SUMMARY The effects of testosterone on the incorporation of leucine and uridine into the proteins and RNA in vitro and on contractile properties of the levator ani muscle were studied. After long-term preincubation of the muscle with testosterone, incorporation of the labelled precursors into RNA and proteins of the muscle increased by 33 and 78% respectively. No previous treatment in vivo was necessary to show this effect of testosterone. The increase of incorporation of the labelled precursors into RNA and proteins, tested after preincubation with testosterone, was more pronounced after castration, and was 61 and 134% respectively. The incorporation of labelled precursors into RNA and proteins of the diaphragm muscle of the rat was not affected by preincubation with testosterone in vitro. Long-term application of testosterone in vitro resulted in a decrease of contraction time and latency period of the levator ani muscle.

EFFECT OF TESTOSTERONEPHENYLPROPIONATE AND 19-NORTESTOSTERONEPHENYLPROPIONATE ON THE SEMINAL VESICLES, THE LEVATOR ANI MUSCLE AND THE MAMMARY GLANDS OF CASTRATED MALE RATS

1962 ◽  
Vol 39 (4) ◽  
pp. 584-598 ◽  
Author(s):  
K. Ahrén ◽  
A. Arvill ◽  
Å. Hjalmarson
Keyword(s):  
Levator Ani ◽  
Mammary Glands ◽  
Seminal Vesicles ◽  
Male Rats ◽  
Weight Increase ◽  
Levator Ani Muscle ◽  
List Type ◽  
Minimal Weight ◽  
Alveolar Development ◽  
Marked Weight

ABSTRACT The response of the seminal vesicles, the levator ani muscle and the mammary glands to testosteronephenylpropionate (TPP) and 19-nortestosteronephenylpropionate (19-norTPP) was studied in castrated male rats. The development of these structures was compared with that found in male rats with intact testes. The main results were as follows: 1) Daily injections of 0.01 mg of TPP produced slight weight increase in the seminal vesicles and levator ani muscle and stimulated a slight but obvious lobule-alveolar development in the mammary glands. The same dose of 19-norTPP produced only a minimal weight increase in the seminal vesicles but produced an obvious development of the levator ani muscle and the mammary glands. 2) Daily injections of 0.05 mg of TPP caused a marked weight increase in the seminal vesicles and levator ani muscle and produced a marked lobule-alveolar development in the mammary glands. The same dose of 19-norTPP produced only a slight weight increase in the seminal vesicles but brought about a marked development of the levator ani muscle and the mammary glands. 3) Daily injections of 0.5 mg of 19-norTPP caused a marked development of the seminal vesicles comparable to that found in rats with intact testes. The levator ani muscle and the mammary glands after this treatment were, however, much more stimulated than in rats with intact testes. These results indicate 1) that the ratio between the effects of these compounds on the seminal vesicles and on the Ievator ani muscle depends on the dose-level and 2) that the development of the mammary glands is correlated more to the growth of the Ievator ani muscle than to the development of the seminal vesicles.

EFFECTS OF TESTOSTERONE ON THE METABOLISM OF THE ISOLATED LEVATOR ANI MUSCLE OF THE RAT

1967 ◽  
Vol 56 (3_Suppl) ◽  
pp. S1-S14 ◽  
Author(s):  
A. Arvill
Keyword(s):  
Incubation Period ◽  
Protein Fraction ◽  
Muscle Protein ◽  
Levator Ani ◽  
Male Rats ◽  
Levator Ani Muscle ◽  
Intracellular Accumulation ◽  
Immature Male

ABSTRACT The effects of restosterone were studied on the rate of intracellular accumulation of D-xylose-14C, AIB-14C, glycine-3H, valine-14C and leucine-14C in the isolated levator ani muscle of immature male rats. The effects of testosterone were also investigated on the incorporation of glycine-3H and leucine-14C into the muscle protein as well as on the incorporation of adenine-14C in muscle RNA. The testosterone stimulated AIB transport is described by the formulation of Michaelis-Menten and approximate values of Km and Vmax are calculated for this transport in the levator ani muscle. Testosterone stimulated the intracellular accumulation of D-xylose-14C, AIB-14C and glycine-3H, but only when injected to the rats at least six hours before the in vitro experiment. No effect could be observed when testosterone was added to the incubation media. Testosterone also stimulated the incorporation of glycine-3H and leucine-14C into the muscle protein. When puromycin was added to the medium no radioactivity was found in the protein fraction. Stimulating effects during the incubation period, of previously injected testosterone, were nevertheless observed on the intracellular accumulation of AIB-14C, glycine-3H and valine-14C. Testosterone also stimulated the incorporation of adenine-14C into the muscle RNA. No effects on these parameters were ever observed in the diaphragm muscles of the same rats.

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