Biomedical subjects

Q Gong

Publications and source records attributed to Q Gong.

At least 55 records · Page 3Linked to original sources

PubMed1998

Properties of HERG channels stably expressed in HEK 293 cells studied at physiological temperature.

We have established stably transfected HEK 293 cell lines expressing high levels of functional human ether-a go-go-related gene (HERG) channels. We used these cells to study biochemical characteristics of HERG protein, and to study electrophysiological and pharmacological properties of HERG channel current at 35 degrees C. HERG-transfected cells expressed an mRNA band at 4.0 kb. Western blot analysis showed two protein bands (155 and 135 kDa) slightly larger than the predicted molecular mass (127 kDa). Treatment with N-glycosidase F converted both bands to smaller molecular mass, suggesting that both are glycosylated, but at different levels. HERG current activated at voltages positive to -50 mV, maximum current was reached with depolarizing steps to -10 mV, and the current amplitude declined at more positive voltages, similar to HERG channel current expressed in other heterologous systems. Current density at 35 degrees C, compared with 23 degrees C, was increased by more than twofold to a maximum of 53.4 +/- 6.5 pA/pF. Activation, inactivation, recovery from inactivation, and deactivation kinetics were rapid at 35 degrees C, and more closely resemble values reported for the rapidly activating delayed rectifier K+ current (I(Kr)) at physiological temperatures. HERG channels were highly selective for K+. When we used an action potential clamp technique, HERG current activation began shortly after the upstroke of the action potential waveform. HERG current increased during repolarization to reach a maximum amplitude during phases 2 and 3 of the cardiac action potential. HERG contributed current throughout the return of the membrane to the resting potential, and deactivation of HERG current could participate in phase 4 depolarization. HERG current was blocked by low concentrations of E-4031 (IC50 7.7 nM), a value close to that reported for I(Kr) in native cardiac myocytes. Our data support the postulate that HERG encodes a major constituent of I(Kr) and suggest that at physiological temperatures HERG contributes current throughout most of the action potential and into the postrepolarization period.

Action Potentials↗

PubMed1998

[Mechanism and treatment of malignant glaucoma].

OBJECTIVE: To investigate the possible mechanism and the treatment of malignant glaucoma. METHODS: The onsets of 34 eyes in 23 cases of malignant glaucoma were analyzed and the patients were treated with medicine, lens extraction, vitrectomy, anterior chamber plastic operation or combined procedures. RESULTS: The effective rate in medical treatment was 35.3%. Combined procedures (lens extraction associated with vitrectomy and anterior chamber plastic operation) were all successful. CONCLUSIONS: The possible mechanism of malignant glaucoma may be concerned with the formation of membranoid substance among vitreous face, ciliary body and posterior iris. Those who failured to medical therapy and were treated with combined operations may have good effect.

Adolescent↗

PubMed1998

[The processing of Gekko gecko and the pharmacological action of its different parts].

Both pharmacologic and toxic experiments are made on different parts of Gekko gecko Linnaeus. The results show that Gekko gecko Linnaeus' heads and its feet have obvious pharmacological action without any toxic or side effects, which provides a sound basis for the increase in its clinical utilization and the expansion of its medicinal parts as well as guarantee of safety and effectiveness after taking the medicine.

Adjuvants, Immunologic↗

PubMed1997

Effect of thiols on cadmium-induced expression of metallothionein and other oxidant stress genes in rat lung epithelial cells.

This study examined cadmium-induced alterations in metallothionein-1 (MT), glutathione-S-transferase Ya (GST), and heme oxygenase-1 (HO) gene expression in an adult rat lung epithelial cell line. Elevations in MT mRNA and HO mRNA occurred as early as 1 h after exposure to a sub-toxic concentration of CdCl(2) (10 microM) whereas GST expression did not increase significantly until 4 h after Cd addition. At t = 8 h, levels of GST, MT, and HO mRNA were elevated 9-fold, 27-fold, and 44-fold, respectively, over basal expression. By 24 h, MT expression was almost back to baseline levels. GST mRNA and HO mRNA were also reduced, compared to 8 h, but to a lesser extent than MT expression. The MT gene was more responsive to low Cd concentrations (5 microM) than the genes for HO or GST whereas HO was induced more than the others at higher Cd doses (10-20 microM). Pro-oxidant conditions play a role in Cd-induced gene expression, as suggested by the rapid decline (15-30 min) in glutathione (GSH), amounting to 25-30% of baseline, that occurred after exposure to 10 microM CdCl(2). This was followed by resynthesis of GSH to a concentration higher than the initial. Depleting GSH by treatment of cells with buthionine sulfoximine (BSO) enhanced Cd-induced expression of MT, GST, and HO whereas thiol supplementation, by treatment with N-acetyl cysteine (NAC), had an attenuating effect. BSO and NAC pretreatment had no effect on basal gene expression or Cd uptake. In summary, this study has shown that: (1) Cd increases MT, GST, and HO gene expression in a time- and dose-dependent fashion: (2) MT gene expression appears to be most sensitive to Cd whereas the HO gene is most inducible at higher Cd concentrations; (3) Cd-induced expression is enhanced by GSH depletion and suppressed by thiol supplementation.

Acetylcysteine↗

PubMed1997

Restoration of thymocyte development and function in zap-70-/- mice by the Syk protein tyrosine kinase.

The Syk family of protein tyrosine kinases, consisting of ZAP-70 and Syk, associate with the pre- and alphabeta T cell antigen receptors (TCRs) and undergo tyrosine phosphorylation and activation following receptor engagement. Thymocyte development in zap-70-/- mice is blocked at the CD4+CD8+ TCR(lo) stage. The presence of Syk in the thymus has raised the possibility that Syk may be able to mediate TCR function. To determine if Syk can play a role in thymocyte development, we generated zap-70-/- mice expressing a human syk cDNA. Syk expression restored both thymocyte development and function. In addition, Syk function required the CD45 transmembrane protein tyrosine phosphatase. Hence, ZAP-70 and Syk can play overlapping functions and exhibit similar regulatory mechanisms in mediating alphabeta T cell development.

Animals↗

PubMed1997

Blockage of the HERG human cardiac K+ channel by the gastrointestinal prokinetic agent cisapride.

Cisapride, a gastrointestinal prokinetic agent, is known to cause long Q-T syndrome and ventricular arrhythmias. The cellular mechanism is not known. The human ether-á-go-go-related gene (HERG), which encodes the rapidly activating delayed rectifier K+ current and is important in cardiac repolarization, may serve as a target for the action of cisapride. We tested the hypothesis that cisapride blocks HERG. The whole cell patch-clamp recording technique was used to study HERG channels stably expressed heterologously in HEK293 cells. Under voltage-clamp conditions, cisapride block of HERG is dose dependent with a half-maximal inhibitory concentration of 6.5 nM at 22 degrees C (n = 25 cells). Currents rapidly recovered with drug washout. The onset of block by cisapride required channel activation indicative of open or inactivated state blockage. Block of HERG with cisapride after channel activation was voltage dependent. At -20 mV, 10 nM cisapride reduced HERG tail-current amplitude by 5%, whereas, at + 20 mV, the tail-current amplitude was reduced by 45% (n = 4 cells). At -20 and + 20 mV, 100 nM cisapride reduced tail-current amplitude by 66 and 90%, respectively. We conclude that cisapride is a potent blocker of HERG channels expressed in HEK293 cells. This effect may account for the clinical occurrence of Q-T prolongation and ventricular arrhythmias observed with cisapride.

Cation Transport Proteins↗

PubMed1996

Pulmonary metallothionein expression in rats following single and repeated exposure to cadmium aerosols.

This study examined the expression of metallothionein (MT) isoforms in the lungs of Lewis rats exposed to Cadmium (Cd) aerosols. With the use of isoform-specific oligonucleotide probes and Northern hybridization analysis, we demonstrated that a dramatic, rapid, and coordinate increase occurred in pulmonary MT-1 mRNA and MT-2 mRNA following Cd inhalation exposure. MT mRNAs levels reached a maximum at 2 h post-exposure and remained above control levels at 96 h after exposure. A considerable lag between the time of maximal elevations in MT mRNAs and MT protein accumulation was observed and suggested that regulatory mechanisms in addition to transcriptional control could be involved. MT expression (protein and mRNA) and Cd lung burden were directly related to aerosol Cd concentration. In situ hybridization and immunohistochemistry studies showed good correlation between the localization of MT protein and MT mRNAs. However, staining for MT protein and MT mRNA was not uniformly distributed in the lung. MT was particularly prominent within the alveolar compartment. Even within this area, however, heterogeneity of MT expression was evident. Experiments were subsequently conducted to determine whether prior exposure to Cd modulates the transcriptional activity of MT genes such that there is a greater elevation in gene expression upon reexposure to Cd. Surprisingly, animals pretreated with Cd exhibited a smaller incremental increase in MT mRNA levels in response to subsequent Cd exposure than controls with no prior treatment. Moreover, MT mRNA levels were elevated to a similar extent regardless of whether animals were exposed to Cd aerosols for 1, 2, or 3 weeks (3 h/day; 5 days/week). MT protein and lung Cd burden, on the other hand, exhibited an increasing linear trend as a function of exposure number. In summary, this study has demonstrated that: (1) the lung responds to Cd inhalation exposure by increasing MT mRNA and MT protein levels; (2) MT expression is prominent within alveolar cells but not all cells are MT positive; and (3) Cd-pretreatment does not increase the transcriptional potential of MT genes when the animal is subsequently reexposed to Cd.

Administration, Inhalation↗

PubMed1996

Modulation of human muscle sodium channels by intracellular fatty acids is dependent on the channel isoform.

Free fatty acids (FFAs), including arachidonic acid (AA), are implicated in the direct and indirect modulation of a spectrum of voltage-gated ion channels. Skeletal muscle sodium channels can be either activated or inhibited by FFA exposure; the response is dependent on both FFA structure and site of exposure. Recombinant human skeletal muscle sodium channels (hSkM1) were transfected into heterologous human renal epithelium HEK293t cells. Cytoplasmic delivery of 5 microM AA augmented the voltage-activated sodium current of hSkM1 channels by 190% (+/-54 S.E., n = 7) over a 20-min period. Similar results were seen with 5 microM oleic acid. Sodium currents in HEK293t cells transfected with human cardiac muscle sodium channels (hH1) were insensitive to AA treatment, and exposure to oleic acid inhibited the hH1 currents over a 20-min period by 29% (+/-13 S.E., n = 5). The increase in hSkM1 current was not accompanied by shifts in voltage dependence of activation, steady-state inactivation, or markedly altered kinetics of inactivation of the macroscopic current. The FFA-induced increase in sodium currents was not dependent on protein kinase C activity. In contrast, both isoforms were reversibly inhibited by external application of unsaturated FFA. Thus, the differential effects of FFA on skeletal muscle sodium channels first noted in cultured muscle cells can be reproduced by expressing recombinant sodium channels in epithelial cells. Although the responses to applied FFAs could be direct or indirect, we suggest that: 1) SkM1 has two classes of response to FFA, one which produces augmentation of macroscopic currents with intracellular FFA, and a second which produces inhibition with extracellular FFA; 2) H1 has only one class of response, which produces inhibition with extracellular FFA. A testable hypothesis is that the presence or absence of each response is due to a specific structure in SkM1 or H1. These specific structures may directly interact with FFA or may interact with intermediate components.

Alkaloids↗

PubMed1996

Increased oxidant resistance of alveolar macrophages isolated from rats repeatedly exposed to cadmium aerosols.

This study investigated potential mechanisms of oxidant resistance in alveolar macrophages (AM) isolated from Lewis rats exposed repeatedly to cadmium aerosols. Macrophages from Cd-adapted animals significantly greater resistance to oxidant-induced cytotoxicity than control cells when challenged with hydrogen peroxide in vitro. Elevations in glutathione peroxidase and glutathione reductase activities were associated with increased oxidant tolerance but catalase activity was unchanged. Metallothionein (MT) expression (protein and mRNA) was dramatically up-regulated in response to in vivo Cd exposure. A study using immunocytochemistry and in situ hybridization techniques revealed significantly heterogeneity in the expression of metallothionein by AMs. The percentage of AMs positive for MT (protein and mRNA) and the degree of MT expression within individual cells increased in response to additional Cd exposures. A putative state of activation was suggested by differences in size and number of inclusion bodies in macrophages from Cd-adapted animals and by secretion of a cytokine with interleukin-1-like characteristics. In summary, AMs from Cd-adapted animals are distinguished from control cells with respect to: (1) increased oxidant resistance, (2) secretion of cytokines, (3) elevations in enzymes associated with glutathione metabolism, and (4) up-regulation in metallothionein expression.

Aerosols↗

PubMed1996

Expression of extracellular matrix molecules and cell surface molecules in the olfactory nerve pathway during early development.

The spatial and temporal expression patterns of several extracellular matrix molecules--laminin and fibronectin and cell surface molecules, neural cell adhesion molecule (NCAM), L1, tenascin, chondroitin sulfate proteoglycan, and peanut agglutinin (PNA) binding sites--were investigated during early olfactory nerve development. NCAM and L1 have similar patterns: They are expressed in the olfactory nerve and on the olfactory receptor neurons (ORNs) commencing with the earliest olfactory axon outgrowth (E12-E15). Their expression patterns suggest that both NCAM and L1 are associated with extension and fasciculation of olfactory axons. A comparison of L1 and olfactory marker protein suggests that L1 is expressed predominantly on immature ORNs. Laminin has an unique punctate staining pattern in the developing olfactory pathway as early as E12. These laminin puncta might play a role in olfactory neurite outgrowth and guidance. At E14, when pioneer olfactory axons enter the brain, the laminin-positive meninges on the surface of the olfactory bulb primordium break down but remain intact in the rest of the telencephalon. This suggests a functional interaction between the olfactory axons and the glial-pial barrier. Fibronectin staining is diffuse throughout the cranial mesenchyme but is absent from the olfactory nerve pathway. No specific patterns of tenascin or chondroitin sulfate, were observed during early olfactory development. PNA binding sites were associated with olfactory axon fasciculation. The expression of several extracellular matrix molecules and cell surface molecules is spatially and temporally regulated in the developing olfactory system. These molecules, thus, may play functional roles in olfactory axon outgrowth, fasciculation, and/or guidance.

Animals↗

PubMed1996

Olfactory epithelial organotypic slice cultures: a useful tool for investigating olfactory neural development.

An in vitro slice culture was established for investigating olfactory neural development. The olfactory epithelium was dissected from embryonic day 13 rats; 400 microns slices were cultured for 5 days in serum-free medium on Millicell-CM membranes coated with different substrates. The slices were grown in the absence of their appropriate target, the olfactory bulb, or CNS derived glia. The cultures mimic many features of in vivo development. Cells in the olfactory epithelium slices differentiate into neurons that express olfactory marker protein (OMP). OMP-positive cells have the characteristic morphology of olfactory receptor neurons: a short dendrite and a single thin axon. The slices support robust axon outgrowth. In single-label experiments, many axons expressed neural specific tubulin, growth-associated protein 43 and OMP. Axons appeared to grow equally well on membranes coated with type I rat tail collagen, laminin or fibronectin. The cultures exhibit organotypic polarity with an apical side rich in olfactory neurons and a basal side supporting axon outgrowth. Numerous cells migrate out of the slices, of which a small minority was identified as neurons based on the expression of neural specific tubulin and HuD, a nuclear antigen, expressed exclusively in differentiated neurons. Most of the migrating cells, however, were positive for glial fibrillary acidic protein and S-100, indicating that they are differentiated glia. A subpopulation of these glial cells also expressed low-affinity nerve growth factor receptors, indicating that they are olfactory Schwann cells. Both migrating neurons and glia were frequently associated with axons growing out of the slice. In some cases, axons extended in advance of migrating cells. This suggests that olfactory receptor neurons in organotypic cultures require neither a pre-established glial/neuronal cellular terrain nor any target tissue for successful axon outgrowth. Organotypic olfactory epithelial slice cultures may be useful for investigating cellular and molecular mechanisms that regulate early olfactory development and function.

Animals↗

PubMed1996

The role of prohormone convertases in insulin biosynthesis: evidence for inherited defects in their action in man and experimental animals.

The hormone insulin remains the cornerstone of diabetic therapy since it is required for almost all cases of Type 1 and many cases of Type 2 diabetes. Since the discovery of insulin in 1921, much has been learned about its chemistry, structure and action as well as its production in the beta cell. Insulin is formed through a series of precursors, beginning with preproinsulin, the protein encoded in the insulin gene. These precursors direct the prohormone into the secretory pathway and ultimately into the secretory granules where it is converted into insulin and C-peptide. These products are stored and secreted together in a highly regulated manner in response to glucose and other stimuli. This review focuses on the recently discovered prohormone convertases, PC2 and PC3 (PC1), the enzymes responsible for the endoproteolytic processing of proinsulin to insulin and C-peptide in the beta cell as well as for the selective processing of proglucagon to glucagon in the alpha cell or GLP1 in intestinal L-cells. PC2 and PC3 are calcium-dependent serine proteases related to the bacterial enzyme subtilisin. They cleave selectively at Lys-Arg or Arg-Arg sites in precursors, generating products with C-terminal basic residues that are then removed by carboxypeptidase E, an exopeptidase. All 3 enzymes are expressed mainly in secretory granules of neuroendocrine cells throughout the body and in the brain. Inherited defects affecting the prohormone-processing enzymes have recently been found in association with unusual syndromes of obesity and other metabolic disorders.

Animals↗

PubMed1995

Increased oxidant resistance of alveolar epithelial type II cells. Isolated from rats following repeated exposure to cadmium aerosols.

Alveolar epithelial type II cells (AEIIC) were isolated from male Lewis rats following repeated in vivo cadmium aerosol exposure and were subsequently evaluated for their oxidant resistance in vitro. AEIIC from Cd-adapted animals removed a greater proportion of hydrogen peroxide from the extracellular milieu and incurred less oxidant-induced cytotoxicity than AEIIC from air controls. This altered response to oxidants occurred coincident with changes in cellular resistance factors. A two-fold increase in glutathione peroxidase activity and a 1.5-fold increase in the activities of glutathione reductase and catalase were observed in Cd-adapted AEIIC compared to control cells. These cells also exhibited a dramatic induction of metallothionein (MT), a thiol-rich protein known to scavenge free radicals in vitro. MT concentration increased as a function of exposure number. MT was localized within the nucleus and cytoplasm of AEIIC by immunocytochemical techniques. MT positive cells showed a wide variation of MT content, particularly in the nucleus. The biochemical and physiological features of these AEIIC may explain, in part, why animals pretreated with Cd aerosols develop cross-tolerance to hyperoxia.

Administration, Inhalation↗

PubMed1995

Synergistic effects of growth factors on the regulation of smooth muscle cell scavenger receptor activity.

Rabbit smooth muscle cells (SMC) express types I and II scavenger receptors (ScR) that are up-regulated by platelet secretion products. In the current studies we investigated the effect of growth factors secreted by platelets on ScR activity in rabbit and human SMC. Platelet-derived growth factor (PDGF BB) and transforming growth factor beta 1 (TGF-beta 1) at 10 ng/ml increased ScR activity in rabbit SMC (by approximately 4- and 2-fold, respectively) but not in human SMC. Epidermal growth factor (EGF) or insulin-like growth factor I (IGF-I) alone had little effect on SMC ScR activity. The growth factors had synergistic effects on ScR activity and on types I and II ScR mRNA expression. In rabbit SMC, PDGF BB, EGF, and TGF-beta 1 together stimulated ScR activity 12-fold. In human SMC, EGF and TGF-beta 1, together with either IGF-I or PDGF BB, stimulated receptor activity approximately 7-fold. Growth factor-mediated induction of ScR activity in rabbit and human SMC was blocked by the tyrosine kinase inhibitor tyrphostin 47, whereas the induction of ScR activity in rabbit but not human SMC was blocked by the protein kinase C inhibitor MDL.29,152. Studies using neutralizing antibodies demonstrated that TGF-beta 1 is the predominant factor in in vitro preparations of platelet secretory products which regulates ScR activity. The growth factors that act synergistically in regulating ScR activity in vitro are all present in atherosclerotic lesions, where they are produced by macrophages, endothelial cells, SMC, and platelets. The data suggest that these growth factors may regulate ScR activity in SMC in vivo and contribute to foam cell formation.

Animals↗

PubMed1995

Molecular modeling of the substrate specificity of prohormone convertases SPC2 and SPC3.

In this paper we describe the results of molecular modeling of the structures of the active sites of two subtilisin-like prohormone convertases (SPCs), SPC2 (PC2) and SPC3 (PC1/PC3). These enzymes are members of a recently discovered family of cellular proteases involved in the processing of precursor proteins. Although these proteases all possess catalytic domains similar to the bacterial subtilisins no tertiary structural data from x-ray analysis are yet available. We have shown that despite the high structural homology of the subtilisins and the SPCs, the structure of the loop which lies immediately below the active sites differs due to the presence of a cis-peptide bond (Tyr167-Pro168) in this loop in the subtilisins and its absence in the SPCs. Accordingly, we have proposed a new alignment for the amino acid sequences of the SPCs in this region. Both SPC2 and SPC3 participate in the processing of prohormones at dibasic cleavage sites, typically Lys-Arg or Arg-Arg. To investigate the structural basis of the substrate specificity of these SPCs, we have carried out molecular mechanic calculations of the optimal arrangement and interactions of peptide substrates containing several residues of arginine or lysine, i.e. Arg, Ala-Ala-Ala-Arg, Arg-Ala-Ala-Arg, Arg-Ala-Arg-Arg, Arg-Ala-Lys-Arg, in the putative active sites. Such subtilisin-based modeling has allowed us to identify those negatively charged residues, Asp and Glu, in the S1, S2, and S4 subsites, which can directly interact with basic residues in the substrates via formation of salt bridges and thereby contribute to the substrate selectivity of the SPCs.

Amino Acid Sequence↗

PubMed1995

Interaction of retinoblastoma gene product with transcription factors ATFa and ATF2.

Two highly related transcription factors, activating transcription factors a and 2 (ATFa and ATF2) are able to activate expression in CHO cells of a reporter gene driven by the human transforming growth factor beta 2 gene promoter. This action is dependent on the intact motif CGTCA which is found as part of the cAMP responsive element in a number of promoters and to which both factors can bind in vitro. The retinoblastoma gene product also weakly stimulates expression of this reporter gene but, in combination with the factors, it exhibits a differential action: additive or greater stimulatory effects with ATF2 but strong inhibition of the actions of ATFa. Thus, although both of these two related factors are able to exert the same apparent effect on this promoter, coexpression of the retinoblastoma gene product reveals at least one significant difference in their actions.

Activating Transcription Factor 1↗

PubMed1995

Proteolytic processing mechanisms in the biosynthesis of neuroendocrine peptides: the subtilisin-like proprotein convertases.

The recent discovery of a novel family of precursor processing endoproteases has greatly accelerated progress in understanding the complex mechanisms underlying the maturation of prohormones, neuropeptides, and many other precursor-derived proteins. At least six members of this family have been found thus far in mammalian species, several having alternatively spliced isoforms, and related enzymes have been identified in many invertebrates, including molluscs, insects, nematodes, and coelenterates. The proprotein convertases are all dependent on calcium for activity and all possess highly conserved subtilisin-like domains with the characteristic catalytic triad of this serine protease (ordered Asp, His, and Ser along the polypeptide chain). Two members of this family, PC2 (SPC2) and PC1/PC3 (SPC3), appear to play a preeminent role in neuroendocrine precursor processing. Both convertases are expressed only in the brain and in the extended neuroendocrine system, while another important family member--furin/PACE (SPC1)--is expressed more ubiquitously, in almost all tissues, and at high levels in liver. SPC2 and SPC3 exhibit acidic pH optima and other properties which enhance their activity in the acidic, calcium-enriched environment of the dense-core secretory granules of the regulated pathway in neuroendocrine cells, while furin has a neutral pH optimum and is localized predominantly to the trans Golgi network where it is retained by a C-terminal transmembrane domain. Furin processes a wide variety of precursors in the constitutive pathway, such as those of growth factors, receptors, coagulation factors, and viral glycoproteins. Recent findings on the processing of proopiomelanocortin, proinsulin, proglucagon, and several other neuroendocrine precursors by SPC2 and SPC3 are discussed, along with information on the structure, properties, evolution, developmental expression, and regulation of the convertases. An inherited defect in the fat/fat mouse which affects the processing of proinsulin, and probably also many other prohormones, due to a point mutation in carboxypeptidase E has recently been identified and has begun to provide new insights into the functional integration of the individual processing steps.

Animals↗

PubMed1995

In vivo expression of metallothionein in rat alveolar macrophages and type II epithelial cells following repeated cadmium aerosol exposures.

This study examined the ability of alveolar macrophages and alveolar type II epithelial cells to accumulate cadmium (Cd) and to express metallothionein (MT). Lung cells were isolated from Lewis rats repeatedly exposed to a Cd aerosol (1.6 mg Cd/m3). Intracellular Cd concentration rose following Cd exposure and showed an increasing rend as a function of exposure number. Alveolar macrophages accrued approximately four times more Cd than type II epithelial cells similarly exposed. Macrophages and type II cells responded to the presence of intracellular Cd by increasing MT protein levels. MT concentration was highly correlated with intracellular Cd. Immunocytochemical studies revealed that not all macrophages and type II cells from Cd-exposed animals were immunopositive for MT and that the intensity of immunostaining varied within each cell population. Although a greater percentage of macrophages were immunopositive for MT than type II cells, a greater proportion of type II cells showed moderate and dark MT staining patterns. Oligonucleotide probes, shown to distinguish between MT-1 and MT-2 mRNA isoforms, were used to test for cell-specific differences in MT isoform gene expression. The basal level of MT-1 mRNA was greater in macrophages than in type II cells. Following Cd administration, the level of MT-1 mRNA and MT-2 mRNA increased in each cell class but the response to Cd was three times greater in alveolar macrophages. Neither macrophages nor type II cells expressed MT mRNA isoforms in equal proportions. Macrophages expressed more MT-1 mRNA when exposed to air and more MT-2 mRNA in response to Cd exposure. Type II cells, on the other hand, expressed more MT-2 mRNA than MT-1 mRNA regardless of whether the cells were exposed to air or Cd. In conclusion, the present study has demonstrated that (1) alveolar macrophages and type II cells respond to in vivo Cd exposure by increasing MT protein and mRNA levels; (2) MT expression is greater in macrophages than in type II cells and correlates well with intracellular Cd concentration; and (3) the MT-2 mRNA to MT-1 mRNA ratio is cell and treatment specific.

Aerosols↗