NADPH-d positive neurons of the lizard Podarcis s. sicula oviduct and their relationship to 17beta-estradiol hormone

The distribution of nicotinamide adenine dinucleotide phosphate reduced diaphorase (NADPH-d) containing neurons was examined in the oviduct of the lizard Podarcis s. sicula and the relationship between these neurons and 17beta-estradiol hormone was studied. NADPH-d-histochemistry and indirect immunofluorescence method were applied to cryostat sections. NADPH-d-nerve structures were found throughout the oviduct. Positive neurons were primarily located in the reproductive oviduct, and were more numerous in the intermuscular and circular muscle layers than in the mucosa. The vagina revealed a reactive nerve population denser than elsewhere. The NADPH-d-positive neurons densities and the 17beta-estradiol plasma levels coincided throughout the lizard sexual cycle. In addition, after 17beta-estradiol treatments, non-reproductive lizards showed an increase of NADPH-d neurons. We suppose that nitric oxide (NO) neurons play an estrogen-dependent role in the oviduct muscle motility.     

Triglyceride Uptake in Muscles in Rats

Exogenous lipid is assimilated with different priorities in adipose tissue regions and varies in the fasting and fed conditions. The quantitative role of uptake of lipid in muscle has not been evaluated. In order to examine the uptake in other than adipose tissues, U¹⁴C-oleic acid in sesame oil was administered orally to conscious rats, and lipid label measured after different times in serum, heart, liver, mesenteric, retroperitoneal, inguinal and epididymal fat pads, as well as in red and white parts of gastrocnemius, extensor digitorum longus and soleus muscles. Lipid uptake in total adipose tissue was calculated from dissected adipose tissues plus lipids extracted from the eviscerated, skinned carcass. Lipid uptake in total muscle tissue was estimated from label in dissected muscles plus that in the carcass, assuming similar intracellular lipid contents and radioactivity as that averaged from dissected muscles. Lipid uptake in the liver was calculated from directly extracted lipid. Four hours after lipid administration to fed rats lipid radioactivity in heart and serum was minimal and had essentially disappeared at 8 hours. Liver label declined rapidly from peak values at or before 4 hours. Adipose tissue radioactivity increased gradually up to 16 hours and then decreased. Label in muscles was highest at 4 hours in the red gastrocnemius, and then decreased, while the other muscles showed a constant radioactivity over the observation period (24 hours). Radioactivity expressed per unit muscle mass seemed to be proportional to the oxidative capacity of muscles. In comparisons between fed and fasted rats at 16 hours, when adipose tissue label peaked, liver, individual muscles and carcass did not show any significant differences while adipose tissue label was fivefold higher in fed than fasted rats. The distribution of total measured lipid radioactivity between total adipose tissue, total muscle tissue and liver in fed rats at this time-point was 76. 8, 14. 4 and 8. 8% respectively, and in the fasted state 26. 4, 51. 6 and 22. 0%. These estimations suggest that lipid uptake in the fed state is dominated by adipose tissue, while in the fasted state the lipid uptake is higher in muscles than adipose tissues. It was concluded that uptake of absorbed, exogenous triglyceride in muscle is of significance, particularly in the fasted state. This lipid has a half life of several days. It is suggested that this lipid is oxidized in situ, contributing with a hidden fraction to lipid energy needs, or partially transferred to adipose tissue. Lipid uptake in muscle probably constitutes a significant fraction of assimilated exogenous lipid, particularly in the fasting state.     

Skeletal muscle damage: a study of isotope uptake, enzyme efflux and pain after stepping

We have studied the occurrence of skeletal muscle uptake of 99mtechnetium pyrophosphate (Tc-PYP), creatine kinase (CK) release and muscle pain in normal subjects after exercise. Five subjects stepped on and off a high bench in such a way that one leg stepped up and the other down. Pain only developed in the muscles used for descending: quadriceps, adductors and gluteal muscles of one leg and the calf muscle of the other. A large rise in plasma CK occurred in four subjects but no increased Tc-PYP muscle uptake was seen in the quadriceps. In the four subjects with high CK effluxes, increased isotope uptake was seen in the thigh adductors used when stepping down; in the two subjects with the largest CK effluxes there was extensive uptake into the gluteal muscles. Muscle pain preceded and was not well correlated with either the magnitude of the enzyme release or the amount and distribution of increased muscle isotope uptake. We conclude that delayed onset muscle pain, the cause of which remains unknown, is a poor indicator of muscle damage as indicated by circulating muscle enzymes and muscle isotope uptake. Tc-PYP uptake by skeletal muscle can provide useful information about the localisation and time course of muscle damage.     

Interrelation between ischium,thigh extending muscles and locomotion in some primates

The relative length of the ischium varies according to the species' locomotory behavior, and is useful in estimation of the total relative weight of all the muscles that arise from it. There are three groups of muscles which are involved in thigh extension. Each of these muscles have other function(s) as well. When thigh extending muscles are well developed in a locomotory category or species, it is not only as a result of a response to their function of thigh extension, but also their other functions. In the macaques and guereza, the relative weight of the thigh extending muscles is high due to the massive hamstrings, particularly thebiceps femoris, and the ischium is relatively long. In the chimpanzee, because theadductor magnus muscle is well developed, the relative weight of the thigh extending muscles is high, and the ischium relatively long. In the gibbon, the relative weight of the thigh extending muscles is medium due to the feeble hamstrings and the heavyadductor magnus muscle, and the ischium is of medium relative length. The relative weight of the thigh extending muscles in the slow loris is close to medium with none of the muscles being more dominant than the others, and having ischium of somewhat medium relative length. In the grand and lesser bush babies, the relative weight of the thigh extending muscles and the relative length of the ischium is medium and small respectively, with none of the groups dominating the others, although they, particularly the lesser bush baby, have a massivesemimembranosus. Discussing the ischium in relation with the hamstring muscles alone is not satisfactory, since the ischium serves the thigh extending muscles, includingadductor magnus andquadratus femoris ventralis, attached on it as a lever. A long lever, in these cases the ischium, is an indication of the relatively heavy muscles that originate from it.     

In vitro poly(ADPribosylation) of estrogen-induced proteins from the oviduct of the lizard Podarcis s. sicula Raf.

Poly(ADPribosylation) of nuclear proteins has been investigated in the nuclei from growing oviducts of intact and estrogen-treated spayed females of the lizard Podarcis s. sicula Raf. Isolated nuclei were incubated with [14C] NAD and nuclear proteins extracted in 0.2 m H2SO4. Labeled acid-soluble proteins were analysed by reverse-phase high-performance liquid chromatography (HPLC) and acetic acid-urea gel electrophoresis. The results reported here indicate that the ADPribosylation reaction is involved in modifying besides histone H2b, tissue specific proteins (SNPs and LMG-O). Moreover, comparable results have been obtained from nuclei prepared from the fully active oviduct of intact animals and spayed lizards stimulated with 17 beta-estradiol. It is concluded that the poly-(ADPribosylation) of hormone-induced proteins might play a role in the differentiation of the lizard oviduct.     

Isolation and characterization of binding proteins for retinol from the cytosol, nucleosol and chromatin of the oviduct magnum of laying hens.

Protein fractions that bind retinol were isolated from the cytosol, nucleosol and chromatin of the oviduct magnum of laying hens. The proteins isolated from the three sources showed similar elution profiles on chromatography through Sephadex G-75 and G-50 columns, and comparable mobility during electrophoresis on sodium dodecyl sulphate/polyacrylamide gels. Their molecular weights were calculated to be around 14500. When oviducts from vitamin A-depleted and vitamin A-repleted immature chicks given oestrogen injections for 6 consecutive days were incubated with [3H]retinyl acetate, uptake of the radioactivity in the nuclei of the vitamin A-depleted tissue was severalfold higher than that in the nuclei from the vitamin A-repleted tissue.     

The disappearance of injected prostaglandins from the circulation of adult female australian field crickets,Teleogryllus commodus

During the first 2 h following injection of 0.02 μCi of [¹⁴C]-prostaglandin E₂ into the abdomen of adult virgin female crickets, T. commodus, the concentration of radioactivity in the circulating hemolymph decreases. The reduction is associated with the increase in radioactivity in the Malpighian tubule/hindgut complex, ovaries, fat body, and, to a much smaller extent, ventral nerve cord and flight muscles. The finding that most, but not all, of the radioactivity in the hindgut is located in the contents of the lumen suggests that a high proportion of prostaglandins circulating in the hemolymph of T. commodus is eliminated by the usual excretory pathway. We suggest that the differential uptake of label from the circulating hemolymph by various tissues may be related to possible physiological functions that remain to be discovered.     

The intracellular distribution of labelled lipoproteins in the tissues of rats under physical loading]

A study was made of the influence of a 3 hour physical loading on the intracellular distribution of 125I-labeled very low density lipoprotein (VLDL), low density lipoprotein (LDL), high density lipoprotein (HDL) in the rat tissues after intravenous injection. The uptake of VLDL by liver was decreased, that of LDL was not altered and that uptake of HDL was increased, Redistribution of labeled lipoproteins between the subcellular structures was found. Accumulation of labeled VLDL in cytosol, microsome and large membranes was observed in muscles. The radioactivity level of VLDL was decreased in lysosomes, whereas the level of HDL was increased, Physical loading increased the uptake of HDL in adrenal glands. Accumulation of radioactivity was found in cytosol and small membranes. The data confirm the important role of lipoproteins of cellular metabolism regulation.     

The relationship among ovarian condition, steroid hormones, and estrous behavior in Anolis carolinensis

The lizard Anolis carolinensis ovulates a single egg alternately from each ovary every 14 days. The ovulated egg enters the ipsilateral oviduct, acquires a shell, and is oviposited 18 or 19 days later. Thus, there is only a single egg in one of the oviducts (one-egg condition) during most of the estrous cycle, but there is a 4-5 day period when a mature, shelled egg is in one oviduct while a recently ovulated egg is in the contralateral oviduct (two-egg condition). Analysis of ovarian follicular diameters indicates that the largest follicle in a single ovary exhibits a growth spurt about halfway through its 28-day vitellogenic growth phase. The corpus luteum, once formed from the ovulated follicle, exhibits a linear decrease in weight, becoming fully regressed just before oviposition of the egg in the ipsilateral oviduct. Plasma concentrations of estradiol-17 beta and progesterone are high in two-egg animals. After oviposition, plasma concentrations of progesterone are still high in one-egg animals whereas those of estradiol-17 beta are much lower. These ovarian and hormonal changes during the estrous cycle are discussed in relation to the probable sources of estradiol-17 beta and progesterone, the control of the alternating pattern of ovulation and oviposition, and the role of steroid hormones in female sexual receptivity in this species.     

Estimation of the forces generated by the thigh muscles for transtibial amputee gait

The forces generated by the muscles with origin on the human femur play a major role in transtibial amputee gait, as they are the most effective of the means that the body can use for propulsion. By estimating the forces generated by the thigh muscles of transtibial amputees, and comparing them to the forces generated by the thigh muscles of normal subjects, it is possible to better estimate the energy output needed from prosthetic devices. The purpose of this paper is to obtain the forces generated by the thigh muscles of transtibial amputees and compare these with forces obtained from the same muscles in the case of normal subjects. Two transtibial amputees and four normal subjects similar in size to the amputees were investigated. Level ground walking was chosen as the movement to be studied, since it is a common activity that most amputees engage in. Inverse dynamics and a muscle recruitment algorithm (developed by AnyBody Technology^®) were used for generating the muscle activation patterns and for computing the muscle forces. The muscle forces were estimated as two sums: one for all posterior muscles and one for the anterior muscles, based on the position of the muscles of the thigh relative to the frontal plane of the human body. The results showed that a significantly higher force is generated by the posterior muscles of the amputees during walking, leading to a general increase of the metabolic cost necessary for one step.     

Functional anatomy of the vagina muscles in the adult western conifer seed bug,Leptoglossus occidentalis (Heteroptera: Coreidae), and its implication for the egg laying behaviour in insects

The anatomy of the female reproductive tract and the nerve-evoked contractions of the vagina muscles and their association with the ovipositor in the western conifer seed bug, Leptoglossus occidentalis (Heidemann) are investigated for the first time. The reproductive tract consists of a set of paired telotrophic ovaries, each containing seven ovarioles, located in the anterior lateral regions of the abdomen. Each ovary is attached to a lateral oviduct which spans most of the abdomen to attach to a relatively short common oviduct that joins the vagina near the rear of the animal. The vagina is associated with a pair of bilaterally symmetrical muscles attached at their posterior ends to lateral extensions of sternite VIII, the valvifer of the Type II ovipositor. From this attachment site, the muscles fan out medially and anteriorly to converge along the dorsal midline of the vagina up to the base of the common oviduct. Vagina muscles respond to a single stimulation of their motor nerves by producing a smooth contraction lasting approximately 1 s. With increasing frequencies of stimulation, the muscle contractions summate to create a tetanic response. The muscles are fatigue resistant being able to maintain the same degree of tension for up to 10 min at 10 Hz stimulation. Visual observation shows that other muscles associated with the valves of the ovipositor behave in a similar fashion to that of the vagina muscles from which the tension recordings were obtained. Fatigue-resistant vagina muscles are discussed in relation to copulation, sperm transport and this insect's ability to deposit a series of eggs directly onto the surface of a conifer needle in a manner by which eight or more blunt-ended eggs are packed end-to-end in a single row.     

Uptake of cyclic adenosine-3,5-monophosphate by cultured mammalian cells and frog muscles

Uptake of 3H-cAMP by isolated frog sartorii muscles and cultured mouse 3T3 and 3T6 cells was studied. It was shown that after 1-2 hours of incubation the intracellular level of cAMP in frog muscles reached 10-20% of its concentration in the incubation medium. About 50% of intracellular radioactivity was represented by chromatographically pure cAMP which testifies to the insignificant cAMP metabolism rate. In the experiments with 3T3 and 3T6 cells the influence of possible admixtures and degradation products on 3H-cAMP uptake was revealed. To avoid these influences it is necessary to measure the uptake of cAMP in the presence of theophylline and with abundance of adenosine. In such experimental conditions the intracellular level of cAMP after 1 hour of incubation did not exceed 10% of its extracellular level, which is similar to values obtained on frog muscles. Permeability coefficient of cAMP for membrane of frog muscles and 3T3 and 3T6 cells was about 10(-9)-10(-8) cm X sec-1.     

Preliminary studies on the uptake of labeled mammalian gonadotropins by ovary and oviduct of the green frog, Rana esculenta, in vivo.

In hypophysectomized females, Rana esculenta, a significant uptake of labeled ovine FSH and LH was evident in both the ovary and oviduct. This binding was specific only for the ovary. Ovariectomy did not modify the affinity of the oviduct for these gonadotropins. The possibility of the presence of discrete receptor molecules for FSH and LH in the ovary of Rana esculenta is discussed.     

Distal oviduct and genital chamber of eriophyoids (Acariformes,Eriophyoidea): refined terminology and remarks on CLSM technique for studying musculature of mites

The general morphology of cuticle-lined internal genitalia and oviduct is analyzed in intact females of the phytophagous mites, Loboquintus subsquamatus and Trisetacus cf bagdasariani (Acari: Eriophyoidea) using tetramethylrhodamine B isothiocyanate-phalloidin, three anaesthetics (magnesium sulphate, lidocaine and CO₂-enriched water) and confocal laser scanning microscopy (CLSM). This is the first protocol adopted for CLSM studying musculature of mites. Revision of the previous terminology of eriophyoid internal genitalia from Nuzzaci and Alberti (Eriophyoid mites: their biology, natural enemies and control. World crop pests 6. Elsevier, Amsterdam, pp 101–150, 1996) resulted in the refinement of the terms “distal oviduct”, “genital chamber” and “spermatheca”. Relative position of the elements of cuticle-lined internal genitalia is discussed and a generalized 3D model and animation (available on-line as supplementary material) of eriophyoid genital apparatus are provided. The wall of eriophyoid oviduct contains strong longitudinal muscles attached to the cuticle genital chamber with folded walls. When the egg is being extruded by contraction of the oviduct muscles, it forms lobes corresponding to the internal topography of the oviduct and genital chamber; these lobes invaginate inward from the gonopore, resulting in the “flower-shaped” figures rarely observed in slide-mounted mites. Gnathosomal muscles (cheliceral muscles and extrinsic muscles of palps) and opisthosomal muscles D₁ of Loboquintus mites are attached to the three posterior depressions near the rear prodorsal shield margin. Prospects of CLSM approach for studying different aspects of mite morphology are briefly discussed.     

Functional anatomy of vagina muscles in the blood-feeding insect,Rhodnius prolixus

The physiology of the muscles associated with the vagina in the blood-feeding insect, Rhodnius prolixus Stal, was investigated with the use of Methylene Blue staining to visualize the anatomy, and a micro force transducer to record spontaneous and neurally-evoked contractions. The vagina is associated with a dorsal muscle and a set of paired lateral muscles. The dorsal muscle extends from the base of the common oviduct to apodemes located laterally on sternite VIII, the first genital segment. The lateral muscles extend from a medially-located apodeme on the posterior edge of sternite VI around each side of the common oviduct to travel posteriorly along the side of the vagina before inserting laterally on apodemes on sternite VIII. The vagina muscles display spontaneous and neurally-evoked contractions that are prolonged but transient. The response to evoked contractions shows that the muscles are innervated by both excitatory and inhibitory motor axons. The degree of tension generated by evoked contractions is dependent on the frequency of stimulation with maximal tension being generated at 20–30 Hz. This tension, which often exceeds 400 mg, is transient and returns to a baseline within 1 to 2 min during continuous stimulation. These results, which are the first to describe this chamber in this well-studied insect, are discussed with respect to the act of egg laying.     

A note on meat quality traits of pheasants (<Emphasis Type="Italic">Phasianus colchicus</Emphasis>)

Meat yield, proximate composition, pH and drip loss of breast and thigh muscles were studied in 29 hunted male and 32 slaughtered female pheasants. In the breast muscles of additional 14 hunted male pheasants, colour, cooking loss and shear force were measured. Weight of muscles of hunted male pheasants was higher than that of slaughtered females, but the percentages of breast and thigh muscles relative to the dressed carcass did not differ. Average protein concentrations in lean muscles were above 22%, and average fat was below 1.2%. In breast muscles, pH₂₄ was lower than in thigh (ca. 5.6 vs. 6.0), and, consequently, drip loss was higher (2.2-3.0% vs. 1.0-1.5%). Colour of breast muscles was characterised by L*a*b* values of about 55, 4 and 8-9, respectively. The shear force of breast muscles was about 30 N/cm².     

Muscle oxygen kinetics at onset of intense dynamic exercise in humans

The present study examined the onset and the rate of rise of muscle oxidation during intense exercise in humans and whether oxygen availability limits muscle oxygen uptake in the initial phase of intense exercise. Six subjects performed 3 min of intense one-legged knee-extensor exercise [65.3 +/- 3.7 (means +/- SE) W]. The femoral arteriovenous blood mean transit time (MTT) and time from femoral artery to muscle microcirculation was determined to allow for an examination of the oxygen uptake at capillary level. MTT was 15.3 +/- 1.8 s immediately before exercise, 10.4 +/- 0.7 s after 6 s of exercise, and 4.7 +/- 0.5 s at the end of exercise. Arterial venous O(2) difference (a-v(diff) O(2)) of 18 +/- 5 ml/l before the exercise was unchanged after 2 s, but it increased (P < 0.05) after 6 s of exercise to 43 +/- 10 ml/l and reached 146 +/- 4 ml/l at the end of exercise. Thigh oxygen uptake increased (P < 0.05) from 32 +/- 8 to 102 +/- 28 ml/min after 6 s of exercise and to 789 +/- 88 ml/min at the end of exercise. The time to reach half-peak a-v(diff) O(2) and thigh oxygen uptake was 13 +/- 2 and 25 +/- 3 s, respectively. The difference between thigh oxygen delivery (blood flow x arterial oxygen content) and thigh oxygen uptake increased (P < 0.05) after 6 s and returned to preexercise level after 14 s. The present data suggest that, at the onset of exercise, oxygen uptake of the exercising muscles increases after a delay of only a few seconds, and oxygen extraction peaks after approximately 50 s of exercise. The limited oxygen utilization in the initial phase of intense exercise is not caused by insufficient oxygen availability.     

Electrical stimulation of human lower extremities enhances energy consumption, carbohydrate oxidation, and whole body glucose uptake.

Our laboratory has recently demonstrated that low-frequency electrical stimulation (ES) of quadriceps muscles alone significantly enhanced glucose disposal rate (GDR) during euglycemic clamp (Hamada T, Sasaki H, Hayashi T, Moritani T, and Nakao K. J Appl Physiol 94: 2107-2112, 2003). The present study is further follow-up to examine the acute metabolic effects of ES to lower extremities compared with voluntary cycle exercise (VE) at identical intensity. In eight male subjects lying in the supine position, both lower leg (tibialis anterior and triceps surae) and thigh (quadriceps and hamstrings) muscles were sequentially stimulated to cocontract in an isometric manner at 20 Hz with a 1-s on-off duty cycle for 20 min. Despite small elevation of oxygen uptake by 7.3 +/- 0.3 ml x kg(-1) x min(-1) during ES, the blood lactate concentration was significantly increased by 3.2 +/- 0.3 mmol/l in initial period (5 min) after the onset of the ES (P < 0.01), whereas VE showed no such changes at identical oxygen uptake (7.5 +/- 0.3 ml x kg(-1) x min(-1)). ES also induced enhanced whole body carbohydrate oxidation as shown by the significantly higher respiratory gas exchange ratio than with VE (P < 0.01). These data indicated increased anaerobic glycolysis by ES. Furthermore, whole body glucose uptake determined by GDR during euglycemic clamp demonstrated a significant increase during and after the cessation of ES for at least 90 min (P < 0.01). This post-ES effect was significantly greater than that of the post-VE period (P < 0.01). These results suggest that ES can substantially enhance energy consumption, carbohydrate oxidation, and whole body glucose uptake at low intensity of exercise. Percutaneous ES may become a therapeutic utility to enhance glucose metabolism in humans.     

Morphology of gonoducts and male genital papilla, in the bluehead wrasse: implications and correlates on the control of gamete release

In the coral reef fish Thalassoma bifasciatum , males vary the number of sperm they release in successive spawnings with individual females in accordance with female body and clutch size. The morphology and histological structure of the male genital papilla, sperm duct, oviduct and surrounding musculature were examined in an effort to elucidate the mechanism permitting control of the number of gametes released during mating. In males, urinary and genital ducts pass separately through a common urogenital papilla and are associated with a striated sphincter muscle and a pair of thin, smooth ligament muscles arising from the first proximal anal fin radial and passing laterally around the sperm duct and oviduct. Within the papilla, the sperm duct resembles a narrow funnel whose inner walls contain longitudinal folds or septa protruding into the lumen of the duct. Dorsal to the papilla, the sperm duct enlarges and is divided into numerous, open chambers by irregular, longitudinal trabeculae. The wall of the duct and the trabeculae contain flat epithelium, smooth muscle and loose connective tissue. In females, the oviduct contains no trabeculae and is not divided into chambers. The ligament muscles are more thoroughly embedded in the sphincter muscle of the rectum than in males. Some ways in which these structures might control gamete release are suggested.