Effect of vitamin E deprivation and exercise training on induction of HSP70

Kelly, D. A., P. M. Tiidus, M. E. Houston, and E. G. Noble.Effect of vitamin E deprivation and exercise training on inductionof HSP70. J. Appl. Physiol. 81(6):2379-2385, 1996.To investigate the effects of dietary vitamin Edeprivation and chronic exercise on the relative content of selectedisoforms of the heat-shock protein 70 (HSP70) family in rat hindlimbmuscle, vitamin E was withheld for 16 wk from female rats thatunderwent treadmill run training during the final 8 wk. As indicated byincreased (P < 0.05) content of thestress-inducible isoform (HSP72), training did stress the exercisingmuscles. However, vitamin E deficiency did not alter HSP72 content innontrained rats and was associated with a lesser induction(P < 0.01) in some muscles oftrained animals. The constitutive isoform, which exhibited similarlevels in muscles of varying fiber types, was demonstrated to belargely refractory to exercise, with an equivocal response to vitamin Edeprivation. HSP72 content was correlated to type I myosin heavy chain(MHC-I) content but only in muscles of sedentary normal-diet rats.After training, HSP72 content in a muscle essentially devoid of MHC-I(superficial vastus lateralis) reached levels comparable to those in amuscle high in MHC-I (soleus).

     

Creatine transporter protein content, localization, and gene expression in rat skeletal muscle

The present study examined the geneexpression and cellular localization of the creatine transporter(CreaT) protein in rat skeletal muscle. Soleus (SOL) and red (RG) andwhite gastrocnemius (WG) muscles were analyzed for CreaT mRNA, CreaTprotein, and total creatine (TCr) content. Cellular location of theCreaT protein was visualized with immunohistochemical analysis ofmuscle cross sections. TCr was higher (P  0.05) in WGthan in both RG and SOL, and was higher in RG than in SOL. Total CreaTprotein content was greater (P  0.05) in SOL and RGthan in WG. Two bands (55 and 70 kDa) of the CreaT protein were foundin all muscle types. Both the 55-kDa (CreaT-55) and the 70-kDa(CreaT-70) bands were present in greater (P  0.05)amounts in SOL and RG than in WG. SOL and RG had a greater amount(P  0.05) of CreaT-55 than CreaT-70. Immunohistochemical analysis revealed that the CreaT was mainly associated with the sarcolemmal membrane in all muscle types. CreaTmRNA expression per microgram of total RNA was similar across the threemuscle types. These data indicate that rat SOL and RG have an enhancedpotential to transport Cr compared with WG, despite a higher TCr in the latter.

     

Intracellular signaling leads to the hypertrophic effect of neuropeptide Y

Signal transduction pathways involved in the hypertrophic effectof neuropeptide Y (NPY) were investigated in adult cardiomyocytes. Reduction of transforming growth factor- activity inserum-supplemented media abolished the induction of hypertrophicresponsiveness to NPY. In responsive cells, NPY (100 nM) increasedprotein synthesis, determined as incorporation of[¹⁴C]phenylalanine, by35 ± 15% (P < 0.05, n = 16 cultures). In these cells, NPYactivated pertussis toxin (PTx)-sensitive G proteins andphosphatidylinositol (PI) 3-kinase. PTx and inhibition of PI 3-kinaseabolished the hypertrophic effect of NPY. NPY also activated proteinkinase C (PKC) and mitogen-activated protein (MAP) kinase. Inhibitionof these two kinases attenuated the induction of creatine kinase(CK)-BB but not the growth response to NPY. In conclusion, NPYstimulates protein synthesis in adult cardiomyocytes via activation ofPTx-sensitive G proteins and PI 3-kinase and it induces the fetal-typeCK-BB via activation of PKC and MAP kinase.

     

Isolated mouse pancreatic beta-cells show cell-specific temporal response pattern

The length of the silent lag time beforeelevation of the cytosolic free Ca²⁺ concentration([Ca²⁺]_i) differs between individualpancreatic -cells. One important question is whether thesedifferences reflect a random phenomenon or whether the length of lagtime is inherent in the individual -cell. We compared the lag times,initial dips, and initial peak heights for[Ca²⁺]_i from two consecutive glucosestimulations (with either 10 or 20 mM glucose) in individualob/ob mouse -cells with the fura 2 technique in amicrofluorimetric system. There was a strong correlation between thelengths of the lag times in each -cell (10 mM glucose:r = 0.94, P < 0.001; 20 mM glucose:r = 0.96, P < 0.001) as well as between theinitial dips in [Ca²⁺]_i (10 mM glucose:r = 0.93, P < 0.001; 20 mM glucose:r = 0.79, P < 0.001) and between theinitial peak heights (10 mM glucose: r = 0.51, P < 0.01; 20 mM glucose: r = 0.77, P < 0.001). These data provide evidence that theresponse pattern, including both the length of the lag time and thedynamics of the subsequent [Ca²⁺]_i, isspecific for the individual -cell.

     

Metabolic effects of low cortisol during exercise in humans

This studyexamined the physiological effect of reduced plasma cortisol (C) duringprolonged exercise in humans. The effects of normal C (NC) werecompared with metyrapone-induced low C (LC) on plasma substrateavailability and the respiratory exchange ratio during 2 h of exerciseat ~60% peak O₂ consumption innine subjects. The C responses were compared with preexercise (Pre) levels and with a rest day (Con). At rest, C was attenuated by ~70%for LC compared with NC. At rest, plasma glucose, lactate, glycerol,-hydroxybutyrate, alanine, branched-chain amino acids, insulin,glucagon, growth hormone, epinephrine, and norepinephrine were similarunder LC and NC (P > 0.05). Duringexercise under NC, plasma C increased compared with Pre, whereas itremained unchanged during LC. During NC, plasma C was elevated at 90 min (compared with Con) and at 120 min (compared with Con and Pre). During exercise, plasma glucose decreased to the same extent and lactate was similar under both conditions, whereas plasma glycerol, -hydroxybutyrate, alanine, and branched-chain amino acids were higher (P < 0.01) under NC. Plasmainsulin declined (P = 0.01) to agreater extent under LC, whereas growth hormone, epinephrine, andnorepinephrine tended to be higher (0.05  P  0.10). Plasma glucagon increasedunder both conditions (P < 0.01).The respiratory exchange ratio did not differ between conditions. Weconclude that, during exercise, 1) Caccelerates lipolysis, ketogenesis, and proteolysis;2) under LC, glucoregulatory hormoneadjustments maintain glucose homeostasis; and3) LC does not alter whole body substrate utilization or the ability to complete 2 h of moderate exercise.

     

Angiogenic growth factor mRNA responses to passive and contraction-induced hyperperfusion in skeletal muscle

It has been proposed that, in skeletal muscle,the angiogenic response to exercise may be signaled by the increase inmuscle blood flow, via biomechanical changes in the microcirculation (increased shear stress and/or wall tension). Toexamine this hypothesis, we compared the change in abundance ofvascular endothelial growth factor (VEGF), basic fibroblast growthfactor (bFGF), and transforming growthfactor-₁(TGF-₁) mRNA in skeletalmuscles of the canine leg after 1 h of pump-controlled high blood flow alone (passive hyperperfusion; protocolA) and electrical stimulation of the femoral andsciatic nerves producing muscle contraction (protocolB). The increase in leg blood flow (5.4- and 5.9-fold change from resting values, respectively) was similar in both groups.Passive hyperperfusion alone did not increase message abundance forVEGF (ratio of mRNA to 18S signals after vs. before hyperperfusion,0.94 ± 0.08) or bFGF (1.08 ± 0.05) but slightly increased thatof TGF-₁ (1.14 ± 0.07;P < 0.03). In contrast, aspreviously found in the rat, electrical stimulation provoked more thana threefold increase in VEGF mRNA abundance (3.40 ± 1.45;P < 0.02). However, electricalstimulation produced no significant changes in either bFGF (1.16 ± 0.13) or TGF-₁ (1.31 ± 0.27). These results suggest that the increased muscle blood flow of exercise does not account for the increased abundance of these angiogenic growth factor mRNA levels in response to acuteexercise. We speculate that other factors, such as localhypoxia, metabolite concentration changes, or mechanical effects ofcontraction per se, may be responsible for the effects of exercise.

     

Tenotomy decreases reporter protein synthesis via the 3'-untranslated region of the beta-myosin heavy chain mRNA

We tested thehypothesis that the -myosin heavy chain (-MHC) 3'-untranslatedregion (UTR) mediates decreased protein expression after tenotomy ofthe rat soleus. We also tested the hypothesis that decreased proteinexpression is the result of RNA-protein interactions within the 3'-UTR.-MHC was chosen for study because of its critical role in thefunction of postural muscles such as soleus. Adult rat soleus muscleswere directly injected with luciferase (LUC) reporter constructscontaining either the -MHC or SV40 3'-UTR. After 48 h oftenotomy, there was no significant effect on LUC expression in the SV403'-UTR group. In the -MHC 3'-UTR group, LUC expression was 37.3 ± 4% (n = 5, P = 0.03) of that in shamcontrols. Gel mobility shift assays showed that a protein factorspecifically interacts with the -MHC 3'-UTR and that tenotomysignificantly increases the level of this interaction (25 ± 7%,n = 5, P = 0.02). Thus the -MHC3'-UTR is directly involved in decreased protein expression that isprobably due to increased RNA-protein binding within the UTR.

     

Cardiovascular adaptations to 10days of cycle exercise

Mier, Constance M., Michael J. Turner, Ali A. Ehsani, andRobert J. Spina. Cardiovascular adaptations to 10 days of cycleexercise. J. Appl. Physiol. 83(6):1900-1906, 1997.We hypothesized that 10 days of training wouldenhance cardiac output (CO) and stroke volume (SV) during peak exerciseand increase the inotropic response to -adrenergic stimulation. Tensubjects [age 26 ± 2 (SE) yr] trained on a cycleergometer for 10 days. At peak exercise, training increasedO₂ uptake, CO, and SV(P < 0.001). Left ventricular (LV)size and function at rest were assessed with two-dimensional echocardiography before (baseline) and after atropine injection (1.0 mg) and during four graded doses of dobutamine. LV end-diastolic diameter increased with training (P < 0.02), whereas LV wall thickness was unchanged. LV contractileperformance was assessed by relating fractional shortening (FS) to theestimated end-systolic wall stress(_ES). Training increased theslope of the FS-_ES relationship (P < 0.05), indicating enhancedsystolic function. The increase in slope correlated with increases inCO (r = 0.71,P < 0.05) and SV(r = 0.70,P < 0.05). The increase in bloodvolume also correlated with increases in CO(r = 0.80, P < 0.01) and SV (r = 0.85, P < 0.004). These datashow that 10 days of training enhance the inotropic response to-adrenergic stimulation, associated with increases in CO and SVduring peak exercise.

     

Effects of the ovarian cycle on sympathetic neural outflow during static exercise

We comparedreflex responses to static handgrip at 30% maximal voluntarycontraction (MVC) in 10 women (mean age 24.1 ± 1.7 yr) during twophases of their ovarian cycle: the menstrual phase (days 1-4) and the follicularphase (days10-12). Changes in muscle sympathetic nerve activity (MSNA; microneurography) in response tostatic exercise were greater during the menstrual compared withfollicular phase (phase effect P = 0.01). Levels of estrogen were less during the menstrual phase(75 ± 5.5 vs. 116 ± 9.6 pg/ml, days 1-4 vs.days 10-12;P = 0.002). Generated tension did not explain differences in MSNA responses (MVC: 29.3 ± 1.3 vs. 28.2 ± 1.5 kg, days 1-4 vs.days 10-12;P = 0.13). In a group of experiments with the use of ³¹P-NMRspectroscopy, no phase effect was observed forH⁺ andH₂PO₄ concentrations(n = 5). During an ischemicrhythmic handgrip paradigm (20% MVC), a phase effect was notobserved for MSNA or H⁺ orH₂PO₄ concentrations,suggesting that blood flow was necessary for the expression of thecycle-related effect. The present studies suggest that, during statichandgrip exercise, MSNA is increased during the menstrual compared withthe follicular phase of the ovarian cycle.

     

Carbohydrate and the cytokine response to 2.5h of running

Nehlsen-Cannarella, S. L., O. R. Fagoaga, D. C. Nieman, D. A. Henson, D. E. Butterworth, R. L. Schmitt, E. M. Bailey, B. J. Warren, A. Utter, and J. M. Davis. Carbohydrate and the cytokineresponse to 2.5 h of running. J. Appl.Physiol. 82(5): 1662-1667, 1997.This randomized,double-blind, placebo-controlled study was designed to determine theinfluence of 6% carbohydrate (C) vs. placebo (P) beverage ingestion oncytokine responses (5 total samples over 9 h) to 2.5 h ofhigh-intensity running (76.7 ± 0.4% maximalO₂ uptake) by 30 experiencedmarathon runners. For interleukin-6 (IL-6), a difference in the patternof change between groups was found, highlighted by a greater increasein P vs. C immediately postrun (753 vs. 421%) and 1.5 h postrun (193 vs. 86%) [F(4,112) = 3.77, P = 0.006]. Forinterleukin-1-receptor antagonist (IL-1ra), a difference in the patternof change between groups was found, highlighted by a greater increasein P vs. C 1.5 h postrun (231 vs. 72%)[F(2,50) = 6.38, P = 0.003]. No significant interaction effects were seen for bioactive IL-6 or IL-1. The immediate postrun plasma glucose concentrations correlated negatively with those of plasma cortisol (r = 0.67, P < 0.001); postrun plasma cortisol (r = 0.70, P < 0.001) and IL-6 levels(r = 0.54, P = 0.003) correlated positively withlevels of IL-1ra. Taken together, the data indicate that carbohydrateingestion attenuates cytokine levels in the inflammatory cascade inresponse to heavy exertion.

     

High glucose and endothelial cell growth: novel effects independent of autocrine TGF-beta 1 and hyperosmolarity

Human endothelial cells wereexposed to 5 mM glucose (control), 25 mM (high) glucose, or osmoticcontrol for 72 h. TGF-1 production, cell growth, death, andcell cycle progression, and the effects of TGF-1 and TGF-neutralization on these parameters were studied. High glucose andhyperosmolarity increased endothelial TGF-1 secretion(P < 0.0001) and bioactivity (P < 0.0001). However, high glucose had a greater effect on reducingendothelial cell number (P < 0.001) and increasingcellular protein content (P < 0.001) than the osmoticcontrol. TGF- antibody only reversed the antiproliferative andhypertrophic effects of high glucose. High glucose altered cell cycleprogression and cyclin-dependent kinase inhibitor expressionindependently of hyperosmolarity. High glucose increased endothelialcell apoptosis (P < 0.01), whereashyperosmolarity induced endothelial cell necrosis (P < 0.001). TGF- antibody did not reverse the apoptotic effectsobserved with high glucose. Exogenous TGF-1 mimicked the increased Sphase delay but not endoreduplication observed with high glucose. High glucose altered endothelial cell growth, apoptosis, and cellcycle progression. These growth effects occurred principally via aTGF-1 autocrine pathway. In contrast, apoptosis andendoreduplication occurred independently of this cytokine and hyperosmolarity.

     

Exercise training improves lusitropy by isoproterenol in papillary muscles from aged rats

Taffet, George E., Lloyd A. Michael, and Charlotte A. Tate.Exercise training improves lusitropy by isoproterenol in papillarymuscles from aged rats. J. Appl.Physiol. 81(4): 1488-1494, 1996.Aging isassociated with a decreased cardiac responsiveness to -adrenergicstimulation. We examined the effect of endurance exercise training ofold Fischer 344 male rats on -adrenergic stimulation of the functionof isolated left ventricular papillary muscle. Three groups wereexamined: sedentary mature (SM; 12-mo old), sedentary old (SO;23-24 mo old), and exercised old (EO; 23-24 mo old) that weretreadmill trained for 4-8 wk. The isometric contractile propertieswere studied at 0.2 Hz and 0.75 mM calcium. Without -adrenergicstimulation, there were no group differences for peak tension, maximumrate of tension development(+dP/dt), or maximum rateof tension dissipation(dP/dt). The time to peak tension was longer (P < 0.05) forboth EO and SO than for SM rats. Half relaxation time(RT_1/2) was prolonged(P < 0.05) for SO compared with SMand EO (which did not differ). The three groups did not differ in the-adrenergic stimulation by isoproterenol of peak tension,dP/dt, time to peak tension, orcontraction duration. The inotropic response(+dP/dt) of SM was greater(P < 0.05) than that in SO or EOrats (which did not differ); however, the lusitropic response(RT_1/2) was lesser(P < 0.05) in SO than in SM or EO rats (which did not differ). Thus exercise training of old rats improved the lusitropic response to isoproterenol without altering theage-associated impairment in inotropic response.

     

Passive sensitization of human airways induces myogenic contractile responses in vitro

Mitchell, R. W., K. F. Rabe, H. Magnussen, and A. R. Leff.Passive sensitization of human airways induces myogenic contractile responses in vitro. J. Appl.Physiol. 83(4): 1276-1281, 1997.We assessedeffects of passive sensitization on human bronchial smooth muscle (BSM)response to mechanical stretching in vitro. Bronchial rings were sham(control) or passively sensitized overnight by using sera from donorsdemonstrating sensitivity to Dermatophagoides farinae and having immunoglobulin E (IgE)concentrations of 2,600 ± 200 U/ml. Tissues were fixedisometrically to force transducers to measure responses to electricalfield stimulation (EFS) and quick stretch (QS). The myogenic responseto QS was normalized to the maximal response to EFS (%EFS). Themyogenic response of sensitized BSM was 47.9 ± 10.9 %EFS to a QSof ~6.5% optimal length (L_o);sham-sensitized tissues had a myogenic response of 13.5 ± 6.4 %EFS(P = 0.012 vs. passively sensitized).A QS of ~13% L_o in sensitizedBSM caused a response of 82.8 ± 20.9 %EFS; sham-sensitized tissuesdeveloped a response of 38.2 ± 17.3 %EFS(P = 0.004). BSM incubated with serumfrom nonallergic donors did not demonstrate increased QS response (4.6 ± 1.4 %EFS, P = not significantvs. tissue exposed to atopic sera). However, tissues incubated in serafrom nonatopic donors supplemented with hapten-specific chimeric IgE(JW8) demonstrated augmented myogenic response to QS of ~6.5% L_o (21.9 ± 6.2 %EFS, P = 0.027 vs. nonatopicsera alone). We demonstrate that passive sensitization of human BSMpreparations causes induction and augmentation of myogenic contractionsto QS; this hyperresponsiveness corresponds to the IgE concentration insensitizing sera.

     

Effect of glucose supplement timing on protein metabolism after resistance training

Roy, B. D., M. A. Tarnopolsky, J. D. MacDougall, J. Fowles,and K. E. Yarasheski. Effect of glucose supplement timing onprotein metabolism after resistance training. J. Appl.Physiol. 82(6): 1882-1888, 1997.We determinedthe effect of the timing of glucose supplementation on fractionalmuscle protein synthetic rate (FSR), urinary urea excretion, and wholebody and myofibrillar protein degradation after resistance exercise.Eight healthy men performed unilateral knee extensor exercise (8 sets/~10 repetitions/~85% of 1 single maximal repetition). Theyreceived a carbohydrate (CHO) supplement (1 g/kg) or placebo (Pl)immediately (t = 0 h) and 1 h(t = +1 h) postexercise. FSR wasdetermined for exercised (Ex) and control (Con) limbs by incrementalL-[1-¹³C]leucineenrichment into the vastus lateralis over ~10 h postexercise. Insulinwas greater (P < 0.01) at 0.5, 0.75, 1.25, 1.5, 1.75, and 2 h, and glucose was greater(P < 0.05) at 0.5 and 0.75 h for CHO compared with Pl condition. FSR was 36.1% greater in the CHO/Ex leg than in the CHO/Con leg(P = not significant) and6.3% greater in the Pl/Ex leg than in the Pl/Con leg(P = not significant). 3-Methylhistidine excretion was lower in the CHO (110.43 ± 3.62 µmol/g creatinine) than Pl condition (120.14 ± 5.82, P < 0.05) as was urinary ureanitrogen (8.60 ± 0.66 vs. 12.28 ± 1.84 g/g creatinine,P < 0.05). This suggests that CHOsupplementation (1 g/kg) immediately and 1 h after resistance exercisecan decrease myofibrillar protein breakdown and urinary urea excretion,resulting in a more positive body protein balance.

     

Effect of leucine metabolite beta -hydroxy-beta -methylbutyrate on muscle metabolism during resistance-exercise training

Nissen, S., R. Sharp, M. Ray, J. A. Rathmacher, D. Rice, J. C. Fuller, Jr., A. S. Connelly, and N. Abumrad. Effect of leucinemetabolite -hydroxy--methylbutyrate on muscle metabolism duringresistance-exercise training. J. Appl.Physiol. 81(5): 2095-2104, 1996.The effects ofdietary supplementation with the leucine metabolite-hydroxy--methylbutyrate (HMB) were studied in two experiments.In study 1, subjects(n = 41) were randomized among threelevels of HMB supplementation (0, 1.5 or 3.0 g HMB/day) and two proteinlevels (normal, 117 g/day, or high, 175 g/day) and weight lifted for1.5 h 3 days/wk for 3 wk. In study 2,subjects (n = 28) were fed either 0 or3.0 g HMB/day and weight lifted for 2-3 h 6 days/wk for 7 wk. Instudy 1, HMB significantly decreased the exercise-induced rise in muscle proteolysis as measured by urine3-methylhistidine during the first 2 wk of exercise (linear decrease,P < 0.04). Plasma creatinephosphokinase was also decreased with HMB supplementation(week 3, linear decrease,P < 0.05). Weight lifted wasincreased by HMB supplementation when compared with the unsupplementedsubjects during each week of the study (linear increase,P < 0.02). In study2, fat-free mass was significantly increased inHMB-supplemented subjects compared with the unsupplemented group at 2 and 4-6 wk of the study (P < 0.05). In conclusion, supplementation with either 1.5 or 3 g HMB/daycan partly prevent exercise-induced proteolysis and/or muscledamage and result in larger gains in muscle function associated withresistance training.

     

Metabolic effects of physical training in ovariectomized and hyperestrogenic rats

This study was undertaken to evaluate theeffects of regular endurance-type exercise on glucose tolerance andglucose-stimulated insulin response (GSIR) in ovariectomized (OVX) ratswith and without estrogen replacement. To do that, OVX Sprague-Dawleyrats were compared with an OVX estradiol-treated group (OVXE2) and asham-operated (Sham) group. Each of these groups was subdivided into asedentary and a treadmill-trained (8 wk) group. Intravenous glucosetolerance tests (0.5 g/kg) were conducted in all rats 48 h afterthe last training session. Plasma levels of 17-estradiol and theuterus weight were significantly (P < 0.05) lower inOVX compared with results in Sham and significantly (P < 0.01) higher in OVXE2 (hyperestrogenic) compared with results inSham. Body weights were significantly (P < 0.01)different among groups, in the following decreasing order: OVX, Sham,and OVXE2. The average daily food intake was significantly(P < 0.01) increased in OVX rats compared with Sham,whereas estradiol treatment diminished this effect (P < 0.01). Exercise training was found to alter none of theabove-mentioned variables in all three experimental conditions. Although the mean integrated area under the glucose and insulin curveswas not affected by OVX, training induced a significant (P < 0.01) reduction in the mean integrated area underthe insulin curve in all three experimental conditions. It is concludedthat the positive effects of physical training on improving GSIR in OVXand hyperestrogenic animals are similar to what has been found in Sham.

     

Immunocompetence of macrophages in rats exposed to Candida albicans infection and stress

The integration of innate andadaptive immune responses is required for efficient control ofCandida albicans. The present work aimed to assess, at thelocal site of the infection, the immunocompetence of macrophages inrats infected intraperitoneally with C. albicans and exposedsimultaneously to stress during 3 days (CaS group). We studied the1) ability to remove and kill C. albicans,2) tumor necrosis factor- (TNF-) release,3) balance of the inducible enzymes NO synthase (iNOS) andarginase, and 4) expression of interleukin (IL)-1 andIL-1 receptor antagonist (ra) mRNA. Compared with only infected animals(Ca group), the number of colony-forming units was significantly higherin CaS rats (P < 0.01), and the macrophagecandidicidal activity was ~2.5-fold lower (P < 0.01). Release of TNF- was diminished in both unstimulated andheat-killed C. albicans restimulated macrophages of the CaSgroup (Ca vs. CaS, P < 0.03 and P < 0.05, respectively). In Ca- and CaS-group rats, the rates for both thearginase activity and the NO synthesis were significantly enhanced.However, the stress exposure downregulated the activity of both enzymes(CaS vs. Ca, P < 0.05). After in vitro restimulation,the IL-1ra/IL-1 ratio was significantly diminished in CaS-group rats(P < 0.05). Our results indicate that a correlationexists between early impairment of macrophage function and stress exposure.

     

Prenatal nicotine affects catecholamine gene expression in newborn rat carotid body and petrosal ganglion

Nicotine exposure modifiesthe expression of catecholamine and opioid neurotransmitter systemsinvolved in attenuation of hypoxic chemosensitivity. We used insitu hybridization histochemistry to determine the effect of prenataland early postnatal nicotine exposure on tyrosine hydroxylase (TH),dopamine -hydroxylase (DH), preproenkephalin (PPE), andD₂-dopamine receptor mRNA levels in the rat carotid bodyand petrosal ganglion during postnatal development. In the carotidbody, nicotine increased TH mRNA expression in animals at 0 and 3 postnatal days (both, P < 0.05 vs. control) withoutaffecting TH mRNA levels at 6 and 15 days. At 15 postnatal days, DHmRNA levels were increased in the carotid body of nicotine-exposed animals. Dopamine D₂-receptor mRNA levels in the carotidbody increased with postnatal age but were unaffected by nicotineexposure. PPE was not expressed in the carotid body at any of the agesstudied in control or treated animals. In the petrosal ganglion,nicotine increased the number of ganglion cells expressing TH mRNA inanimals at 3 days (P < 0.01 vs. control). DH mRNAexpression was not induced nor was PPE mRNA expression increased in thepetrosal ganglion in treated animals. Prenatal nicotine exposureupregulates mRNAs involved in the synthesis of two inhibitoryneuromodulators, dopamine and norepinephrine, in peripheral arterialchemoreceptors, which may contribute to abnormalities incardiorespiratory control observed in nicotine exposed animals.

     

Nerve-responsive troponin I slow promoter does not respond to unloading

We examined theregulation of the troponin I slow (TnIs) promoter during skeletalmuscle unloading-induced protein isoform transition, by using atransgenic mouse line harboring the 4,200 to +12 base pairsregion of the human TnIs promoter. Eighteen female transgenic mice(~30 g body mass) were randomly divided into two groups:weight-bearing (WB) controls (n = 9)and hindlimb unloaded (HU; n = 9). TheHU mice were tail suspended for 7 days. Body mass was unchanged in theWB group but was reduced (6%; P < 0.05) after the HU treatment.Absolute soleus muscle mass (25%) and soleus mass relative tobody mass (16%) were both lower(P < 0.05) in the HU group comparedwith the WB mice. Northern blot analyses indicate that 7 days of HUresult in a 64% decrease (P < 0.05)in the abundance of endogenous TnIs mRNA (µg/mg muscle) in the mousesoleus. Furthermore, there is a trend for the abundance of the fasttroponin I mRNA to be increased (+34%). Analysis of transgenicchloramphenicol acetyltransferase activity in the soleus musclerevealed no difference (P > 0.05)between WB and HU groups. We conclude that additional elements arenecessary for the TnIs gene to respond to an unloading-induced,slow-to-fast isoform transition stimulus.

     

The heat shock response of an antarctic alga is evident at 5°C

A subtidal seaweed collected in antarctic waters, Plocamium cartilagineum (L. Dix.), displayed induction of mRNAs encoding the 70 kDa heat shock protein (HSP70) and the ubiquitin polyprotein (UBI) when incubated at 5°C. Maximal induction of HSP70 mRNA was observed when the alga was incubated at 10°C for 1 h. Incubations at higher temperatures or for longer periods reduced the amount of HSP70 mRNA detected. Incubations at 20°C or greater resulted in cell death. These data indicate that dispite the unusually low temperature of induction, this macrophyte exhibits a heat shock response similar to that of other organisms at temperatures 5 to 10°C above usual growth conditions.