Vascular endothelial growth factor protects post-ganglionic sympathetic neurones from the detrimental effects of hydrogen peroxide by increasing catalase.

AIM: Vascular production of hydrogen peroxide (H(2)O(2)) is implicated in the development and progression of vascular disease. Hydrogen peroxide also promotes neuronal degeneration, which suggests that vascular H(2)O(2) would promote degeneration of perivascular sympathetic nerves. Vascular cells also produce vascular endothelial growth factor (VEGF), which could protect perivascular nerves from the detrimental effects of H(2)O(2) . The aim of this study was to test these hypotheses. METHODS: The effects of H(2)O(2) and VEGF on neuronal survival and noradrenaline uptake were studied in cultures of rat post-ganglionic sympathetic neurones. Western analyses of catalase and growth associated protein 43 were performed and reactive oxygen species (ROS) were measured using the fluorescent indicator 5-(and-6)-chloromethyl-2'7'-dichlorodihydrofluorescein diacetate, acetyl ester. RESULTS: Hydrogen peroxide (30 µm) decreased the survival of post-ganglionic sympathetic neurones (57.8 ± 4.8% of control) and decreased noradrenaline uptake into the neurones (14 ± 6% of control). Hyperglycaemia, which is known to increase H(2)O(2), also decreased survival (31.4 ± 12% of control) and noradrenaline uptake (42 ± 18.4% of control). VEGF reduced the effects of H(2)O(2) (94.3 ± 12% of control) and hyperglycaemia (83.5 ± 23.6% of control) on survival. VEGF increased catalase, a primary determinant of intracellular concentrations of H(2)O(2) , and decreased H(2)O(2) -induced increases in ROS. CONCLUSION: These results indicate that VEGF protects post-ganglionic sympathetic neurones from the detrimental effects of H(2)O(2). Our data suggest that an increase in catalase is the mechanisms underlying this neuroprotection.

PC12 cells stimulate vascular smooth muscle growth.

Sympathetic nerves stimulate vascular growth. The mechanisms underlying this stimulation have not been fully elucidated. PC12 cells and cultures of vascular smooth muscle were used to study sympathetic stimulation of vascular smooth muscle growth. Media conditioned by undifferentiated and differentiated PC12 cells stimulated the growth of vascular smooth muscle (446 +/- 47%). Differentiated PC12 cells produced more growth-stimulatory activity (61.5 +/- 9.6 per 10(6) cells) than undifferentiated PC12 cells (28.5 +/- 8.8 per 10(6) cells). PC12 stimulation of vascular smooth muscle growth was not inhibited by adrenergic receptor antagonists but was reduced by an endothelin antagonist, suramin, and an antibody that neutralized the activity of platelet-derived growth factor. These data suggest that endothelin and platelet-derived growth factor, but not catecholamines, play a role in sympathetic stimulation of vascular smooth muscle growth.

NGF-independent survival of postganglionic sympathetic neurons in neuronal-vascular smooth muscle cocultures.

The present study tests the hypothesis that vascular cells promote the survival of postganglionic sympathetic neurons in the absence of nerve growth factor (NGF). To test this hypothesis, neurons isolated from superior cervical ganglia of 2- to 4-day-old rat pups were grown in the absence of NGF and in the absence and presence of vascular smooth muscle cells (VSM). Neuronal survival was assessed as a function of time in culture. At all time points studied, VSM promoted the survival of the neurons. After 5 days in the absence of NGF, 7 +/- 2% of neurons survived in the absence and 28 +/- 7% survived in the presence of VSM. An endothelin receptor antagonist reduced neuronal survival in cocultures grown in the absence of NGF. These data indicate that VSM produce factors other than NGF that promote the survival of cultured postganglionic sympathetic neurons. The data also indicate that endothelin contributes to this effect and suggest that endothelin as well as other VSM-derived factors may play a role in the development of sympathetic innervation to the vasculature.

Transforming growth factor-beta regulation of endothelin expression in rat vascular cell and organ cultures.

Transforming growth factor (TGF)-beta increases the production of the vasoactive peptide endothelin (ET) in cultures of vascular endothelial cells (EC) and vascular smooth muscle cells (VSMC), but the physiologic or pathologic significance of this regulation has not been determined. The present studies test the hypothesis that when EC and VSMC are in direct contact or close proximity, ET expression is, at least in part, dependent on TGF-beta. The effects of TGF-beta on ET-1 mRNA (Northern analysis and reverse transcription polymerase chain reaction) and peptide (radioimmunoassay) levels were assessed in rat EC and VSMC and vascular organ cultures. TGF-beta2 (1 ng/ml) increased ET-1 mRNA in VSMC and EC plus VSMC cultures and increased ET-1 peptide in EC, VSMC, and EC plus VSMC cultures. TGF-beta2 also increased ET-1 mRNA and peptide in vascular organ cultures. Antibodies that neutralized the activities of TGF-beta1 and TGF-beta2 decreased ET-1 mRNA in EC plus VSMC cultures and in vascular organ cultures. These data indicate that when EC and VSMC are in direct contact or close proximity, TGF-beta increases ET expression and active TGF-beta is present and promotes ET expression. These data suggest that TGF-beta is a determinant of vascular ET expression in vivo, and that TGF-beta regulation of ET expression would affect cardiovascular function in health and disease.

Adrenoceptor-mediated modulation of endothelial-dependent vascular smooth muscle growth.

1. Signals derived from endothelial cells (EC) and the sympathetic nervous system are known to independently modulate the growth of vascular smooth muscle (VSM). It is not known if and how these signals are integrated. The purpose of the present study was to test the hypothesis that activation of adrenoceptors by sympathetic-derived catecholamines modulates EC regulation of VSM growth. 2. The effects of adrenergic agonists on VSM growth were studied in vitro in EC/VSM cocultures. EC stimulated VSM growth in EC/VSM cocultures. Activation of beta-adrenoceptors inhibited this stimulation. EC stimulation of VSM growth was 225+/-31% in the absence and 127+/-27% in the presence of 10 microM isoprenaline. Activation of alpha-adrenoceptors had no effect on EC stimulation of VSM growth in coculture. 3. Isoprenaline did not affect the growth of VSM grown in the absence of EC, suggesting that it did not inhibit EC stimulation of VSM growth by directly inhibiting VSM growth. 4. Isoprenaline did not affect EC production of growth factors, as media conditioned by EC grown in the absence or presence of isoprenaline, stimulated VSM growth to the same extent. Isoprenaline did not alter EC, VSM or EC/VSM production of the VSM growth inhibitor transforming growth factor beta-1 (TGF-beta1). 5. These data provide evidence that catecholamines, via activation of beta-adrenoceptors, can modulate EC-dependent VSM growth, and suggest that the sympathetic nervous system and EC coordinately regulate VSM growth.

VSM growth is stimulated in sympathetic neuron/VSM cocultures: role of TGF-beta2 and endothelin.

Sympathetic nerves are purported to stimulate blood vessel growth. The mechanism(s) underlying this stimulation has not been determined. With use of an in vitro coculture model, the present study tests the hypothesis that sympathetic neurons stimulate the growth of vascular smooth muscle (VSM) and evaluates potential mechanisms mediating this stimulation. Sympathetic neurons isolated from superior cervical ganglia (SCG) stimulated the growth of VSM. Growth of VSM in the presence of SCG (856 +/- 81%) was significantly greater than that in the absence of SCG (626 +/- 66%, P < 0.05). SCG did not stimulate VSM growth in transwell cocultures. An antibody that neutralized the activity of transforming growth factor-beta2 (TGF-beta2) inhibited SCG stimulation of VSM growth in coculture. SCG stimulation of VSM growth was also inhibited by an endothelin A receptor antagonist. These data suggest novel mechanisms for sympathetic modulation of vascular growth that may play a role in the physiological and/or pathological growth of the vasculature.

Endothelin and post-ganglionic sympathetic neurons.

1. The present report documents evidence suggesting that endothelin (ET) is a mediator and modulator of post-ganglionic sympathetic neuronal development. 2. Endothelin is produced by post-ganglionic neurons and by cells adjacent to these neurons. 3. Post-ganglionic sympathetic neurons express functional receptors for ET. 4. Endothelin promotes the survival of cultured post-ganglionic sympathetic neurons and modulates the morphological and biochemical differentiation of these neurons. 5. Endothelin enhances the activity of nerve growth factor and modulates the release of neurotransmitter from post-ganglionic sympathetic nerve terminals.

Ontogeny of local sarcoplasmic reticulum Ca2+ signals in cerebral arteries: Ca2+ sparks as elementary physiological events.

Ca2+ release through ryanodine receptors (RyRs) in the sarcoplasmic reticulum is a key element of excitation-contraction coupling in muscle. In arterial smooth muscle, Ca2+ release through RyRs activates Ca2+-sensitive K+ (KCa) channels to oppose vasoconstriction. Local Ca2+ transients ("Ca2+ sparks"), apparently caused by opening of clustered RyRs, have been observed in smooth and striated muscle. We explored the fundamental issue of whether RyRs generate Ca2+ sparks to regulate arterial smooth muscle tone by examining the function of RyRs during ontogeny of arteries in the brain. In the present study, Ca2+ sparks were measured using the fluorescent Ca2+ indicator fluo-3 combined with laser scanning confocal microscopy. Diameter and arterial wall [Ca2+] measurements obtained from isolated pressurized arteries were also used in this study to provide functional insights. Neonatal arteries (<1 day postnatal), although still proliferative, have the molecular components for excitation-contraction coupling, including functional voltage-dependent Ca2+ channels, RyRs, and KCa channels and also constrict to elevations in intravascular pressure. Despite having functional RyRs, Ca2+ spark frequency in intact neonatal arteries was approximately 1/100 of adult arteries. In marked contrast to adult arteries, neonatal arteries did not respond to inhibitors of RyRs and KCa channels. These results support the hypothesis that RyRs organize during postnatal development to cause Ca2+ sparks, and RyRs must generate Ca2+ sparks to regulate the function of the intact tissue.

Postganglionic sympathetic neurons express endothelin.

Endothelin (ET) is a peptide originally identified as an endothelial-derived vasoconstrictor. It is now recognized that ET is produced by and acts on many other tissues including the brain and spinal cord, where it is believed to modulate neurotransmission. The present studies demonstrate that ET is synthesized by and secreted from postganglionic sympathetic neurons. With the use of Northern analysis, ET-1 mRNA was detected in cultures of sympathetic superior cervical ganglion (SCG) neurons isolated from 3- to 5-day old rat pups. ET-1 and ET-3 peptides were also detected in cultured SCG neurons using immunohistochemistry. ET-1 (50 pg/106 cells) and ET-3 (173 pg/106 cells) were detected by radioimmunoassay of media conditioned by cultured SCG. ET-1 (77 pg/mg protein) and ET-3 (30 pg/mg protein) were also detected by radioimmunoassay of extracts of adult SCG.

The rationale, evolution and clinical application of the self-ligating bracket.

There is ample evidence in the literature that conventional orthodontic mechanics while intended to move teeth efficiently rarely achieve atraumatic remodeling of periodontal tissues. The vascular supply is often interrupted, which in turn affects the oxidative metabolism. Moreover, teeth splinted in groups do not appear to move as efficiently as single teeth. The novel bracket design and treatment regimen described in this report allow teeth to move individually, yet stay within a group. The self-ligating bracket design allows for rapid leveling because teeth drift along the path of least resistance with little or no friction between the bracket and slot of the wire. The hinge mechanism eliminates much of the friction created by the conventional wire or latex ties used to secure the archwire in the bracket slot. Thus, sliding mechanics is achieved in the true sense of the word. This system is capable of increasing the appointment intervals, and possibly reducing the overall treatment time.

In vitro and in vivo vascular actions of basic fibroblast growth factor (bFGF) in normotensive and spontaneously hypertensive rats.

In the cardiovascular system, basic fibroblast growth factor (bFGF) is an important modulator of blood vessel growth and blood pressure and, as such, could contribute to the structural and functional changes that contribute to hypertension. This study evaluated in vitro and in vivo vascular actions of bFGF in normotensive and hypertensive rats. Basic FGF increased the expression of the messenger RNA (mRNA) encoding for the immediate early gene, egr-1, in cultured vascular smooth muscle cells (VSM) isolated from normotensive Wistar-Kyoto (WKY). Sprague-Dawley (SD), and spontaneously hypertensive rats (SHRs). Maximal increases (stimulated/control) in egr-1 mRNA were greater in SHR than in WKY and SD rat VSM (14.57 +/- 1.94 vs. 5.75 +/- 0.35 and 3.84 +/- 0.70). Basic FGF (30 ng/ml) stimulated the growth of WKY (43 +/- 8.4% of growth in 10% serum) and SD (34.6 +/- 6.5%) rat and SHR (75.8 +/- 8.8%) VSM but was most efficacious at stimulating SHR VSM. Radioligand-binding assays indicated no differences in the affinity or number of high-affinity receptors but that the binding of bFGF to low-affinity receptors was slightly but significantly greater in SHR VSM. In vivo, bFGF vasodilated cremaster arterioles in normotensive but not in hypertensive rats. These data suggest that hypertensive animals are more responsive to the growth-stimulatory actions but are less responsive to the vasodilatory actions of bFGF. This altered bFGF function could contribute to the development or maintenance or both of hypertension.

Vascular cell interactions modulate the expression of endothelin-1 and platelet-derived growth factor BB.

Endothelin (ET)-1 and platelet-derived growth factor (PDGF) BB are important regulators of vascular cell growth and contractile function. We tested the hypothesis that interactions between endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) regulate the expression of these two factors. Bovine and rat cocultures of EC-VSMC that allowed cell contact and exchange of soluble mediators were studied. In bovine cocultures, ET-1 and PDGF B transcription decreased. Minimum levels of transcription were 39 +/- 9% of control for ET-1 and 31 +/- 6% of control for PDGF B. In contrast, steady-state mRNA levels transiently increased in coculture; maximum mRNA levels were 579 +/- 248 and 267 +/- 41% of control for ET-1 and PDGF B, respectively. ET-1 and PDGF BB protein levels were decreased in bovine cocultures. The amount of ET-1 in cocultures was 51% less and the amount of PDGF BB was 25% less than that in homotypic cultures. In rat cocultures, ET-1 protein levels were transiently increased compared with homotypic cultures; the maximum difference was 99 +/- 26%. These results indicate that EC-VSMC interactions regulate ET-1 and PDGF BB expression in EC-VSMC cocultures via multiple mechanisms. Furthermore, the interactive regulation appears to be species specific. These data suggest a novel mechanism for the regulation of these two vasoactive factors that may play an important role in cardiovascular physiology and/or pathology.

Nerve growth factor induces transcription of NGFIA through complex regulatory elements that are also sensitive to serum and phorbol 12-myristate 13-acetate.

The expression of NGFIA (also known as egr1, zif268, TIS8, krox24, and d2) is rapidly and transiently increased by nerve growth factor (NGF) in PC12 cells. The 5'-region of this gene includes four serum response elements (SREs), a cAMP-like response element, an AP1-like response element, and an SP1-binding site. From deletion analysis of chloramphenicol acetyltransferase reporter constructs, we have established that the first 106 basepairs 5' of the transcriptional start site are sufficient for induction of NGFIA by NGF in PC12 cells; deletion beyond this point results in dramatically reduced induction of the gene. Using defined mutations in the NGFIA promoter and NGFIA-thymidine kinase hybrid promoters, we have defined three elements (SRE1, SRE2, and AP1-like) in the first 106 basepairs of upstream DNA, each of which contributes to induction of NGFIA by NGF. Cooperation by two of these elements (i.e. the two SREs or one SRE and the AP1-like element) is sufficient to confer transcriptional induction by NGF, but the combination of all three elements increased induction by NGF more effectively than a pair of elements. This suggests that the response of NGFIA to NGF is mediated by a cis-acting sequence that is composed of at least three distinct elements. An oligonucleotide composed of SRE1 and SRE2 that can confer the ability for NGF induction to heterologous promoter constructs complexes with proteins in PC12 cell nuclear extracts, but the protein-DNA complexes do not appear to be altered by NGF treatment, as measured by DNA mobility shift assays. We have also established that the regulatory region of NGFIA that mediates NGF induction also mediates the induction by serum and phorbol 12-myristate 13-acetate, suggesting that multiple signal transduction pathways must converge on these sequences to regulate the expression of this gene.

Rapid fibroblast growth factor-induced increases in protein phosphorylation and ornithine decarboxylase activity: regulation by heparin and comparison to nerve growth factor-induced increases.

Fibroblast growth factors (FGFs), like nerve growth factor (NGF), induce morphological differentiation of PC12 cells. This activity of FGF is regulated by glycosaminoglycans. To further understand the mechanisms of FGF and glycosaminoglycan actions in PC12 cells, we studied the regulation of protein phosphorylation and ornithine decarboxylase (ODC) activity by FGF in the presence and absence of heparin. As with NGF, aFGF and bFGF increased the incorporation of radioactive phosphate into the protein tyrosine hydroxylase (TH). The increase in TH phosphorylation was localized to the tryptic peptide, T3. Both T3 and T1 phosphorylations occur in response to NGF, but there was no evidence that aFGF or bFGF stimulated the phosphorylation of the T1 peptide. This result suggests differential regulation of second messenger systems by NGF and FGF in PC12 cells. Heparin, at a concentration that potentiated aFGF-induced neurite outgrowth 100-fold (100 micrograms/ml), did not alter the ability of aFGF to increase S6 phosphorylation or ODC activity. One milligram per milliliter of heparin, a concentration that inhibited bFGF-induced neurite outgrowth, also inhibited bFGF-induced increases in S6 phosphorylation and ODC activity. These observations suggest (i) that acidic and basic FGF activate a protein kinase, possibly protein kinase C, resulting in the phosphorylation of peptide T3 of TH; (ii) that the FGFs and NGF share some but not all second messenger systems; (iii) that heparin potentiates aFGF actions and inhibits bFGF actions in PC12 cells via distinct mechanisms; (iv) that heparin does not potentiate the neurite outgrowth promoting activity of aFGF by enhancing binding to its PC12 cell surface receptor; and (v) that heparin may coordinately regulate several activities of bFGF (induction of protein phosphorylation, ODC and neurite outgrowth) via a common mechanism, most likely by inhibiting the productive binding of bFGF to its PC12 cell surface receptor.

Nerve growth factor and fibroblast growth factor regulate neurite outgrowth and gene expression in PC12 cells via both protein kinase C- and cAMP-independent mechanisms.

Nerve growth factor (NGF), acidic fibroblast growth factor (aFGF), and basic fibroblast growth factor (bFGF) promote the survival and differentiation of a variety of peripheral and central neurons. The signal transduction mechanisms that mediate the actions of these factors in neuronal cells are not well understood. We examined the effect of a deficiency in protein kinase C (PKC) and/or cAMP second messenger systems on the actions of NGF, aFGF, and bFGF in the pheochromocytoma (PC12) cell line. Activation of PKC was not required for NGF, aFGF, and bFGF to maximally induce ornithine decarboxylase (ODC), transcription of the early response genes, d2 and d5, or neurite outgrowth. In a PC12 cell mutant that is deficient in cAMP responsiveness (A126-1B2), all three growth factors maximally induced the transcription of d5 and neurite outgrowth, but aFGF and bFGF did not induce significant increases in ODC. NGF and aFGF maximally induced the transcription of d2 in A126-1B2 cells, but bFGF-induced d2 transcription was attenuated. NGF, aFGF, and bFGF maximally induced neurite outgrowth and d5 transcription in A126 cells that were made deficient in PKC. The d2 transcriptional response was substantially reduced in cells deficient in both PKC and cAMP responsiveness. These observations lead us to conclude that (a) cAMP- and PKC-dependent events are, at least in part, causally linked to NGF, aFGF, and bFGF induction of both ODC and transcription of d2 and may control functionally redundant pathways; (b) NGF, aFGF, and bFGF can elicit neurite outgrowth and increase transcription of d2 and d5 in PC12 cells via mechanisms that are independent of both PKC and cAMP; (c) NGF, aFGF, and bFGF can induce ODC in the absence of PKC; and (d) aFGF and bFGF require cAMP responsiveness to induce ODC in PC12 cells.

Heparin potentiates the action of acidic fibroblast growth factor by prolonging its biological half-life.

The mechanism(s) by which heparin influences the biological activities of acidic and basic fibroblast growth factors (aFGF and bFGF) is not completely understood. One mechanism by which heparin could alter the biological activities of aFGF and bFGF is by altering their biological half-lives. We investigated the possibility that heparin potentiates aFGF-induced neurite outgrowth from PC12 cells by prolonging its biological half-life. Under conditions where heparin potentiated aFGF-induced neurite outgrowth, we observed that heparin increased the biological half-life of aFGF from 7 to 39 hr. We determined that greater than 25 hr of exposure to active aFGF was required for induction of neurite outgrowth. If aFGF activity was maintained for greater than 25 hr by periodic readdition of factor, heparin no longer potentiated aFGF-induced neurite outgrowth. These observations strongly suggest that heparin potentiates the activity of aFGF by prolonging its biological half-life. The protease inhibitors hirudin, leupeptin, and pepstatin A did not potentiate aFGF-induced neurite outgrowth, indicating that proteases inhibited by these inhibitors are not responsible for the loss of aFGF activity that we observed. However, aprotinin potentiated aFGF neurite-promoting activity approximately sevenfold, indicating that proteases that are inhibited by aprotinin are at least partially responsible for aFGF inactivation. These observations suggest that heparin regulates the activity of aFGF by regulating its proteolytic degradation, thereby regulating its biological half-life.

Sulfated glycosaminoglycans modify growth factor-induced neurite outgrowth in PC12 cells.

Glycosaminoglycans (GAGs), localized on the surfaces of cells and in the basement membrane, modulate the growth and differentiation of many cell types. Recent studies have shown that heparin, a GAG found in mast cells, potentiates the ability of acidic fibroblast growth factor (aFGF) to induce neurite outgrowth in pheochromocytoma (PC12) cells. We examined the effect of a variety of GAGs on aFGF, basic fibroblast growth factor (bFGF), and nerve growth factor (NGF)-induced neurite outgrowth in PC12 cells. The effects observed were dependent upon the specific GAG, the concentration of the GAG, and the growth factor. Heparin potentiated aFGF-induced neurite outgrowth in a concentration-dependent fashion; potentiation increased with increasing heparin concentrations of 0.01-100 micrograms/ml. At concentrations greater than 100 micrograms/ml, heparin potentiation decreased. The maximally active concentration of heparin (100 micrograms/ml) increased the potency of aFGF 102-fold. Increasing concentrations of heparan sulfate, dermatan sulfate, and chondroitin sulfate correlated with increasing aFGF potentiation. The maximally active concentrations of heparan sulfate (100 micrograms/ml), dermatan sulfate (10 mg/ml), and chondroitin sulfate (1 mg/ml) increased the activity of aFGF 11-, 110-, and 11-fold, respectively. Hyaluronic acid did not affect the neurite outgrowth-promoting activity of aFGF. Heparin also altered the activity of bFGF; increasing concentrations of heparin (0.01-1 micrograms/ml) correlated with increased potentiation. At concentrations greater than 1 microgram/ml, heparin concentration was inversely correlated with potentiation. Chondroitin sulfate only increased the percentage of neurite-bearing cells at concentrations greater than 10 micrograms/ml. Maximally active concentrations of heparin (1 microgram/ml) and chondroitin sulfate (1 mg/ml) increased the potency of bFGF 5-fold. The highest concentration of heparan sulfate studied (1 mg/ml) inhibited the activity of bFGF. Dermatan sulfate and hyaluronic acid (0.01-1000 micrograms/ml) had no effect on bFGF activity. Heparan sulfate and chondroitin sulfate showed concentration-dependent potentiation of NGF; maximally active concentrations of heparan sulfate (100 micrograms/ml) and chondroitin sulfate (1 mg/ml) increased the potency of NGF 3-fold, whereas heparin, dermatan sulfate and hyaluronic acid had no effect. None of the GAGs had any effect on PC12 neurite outgrowth when added alone. The specificity of the activity of the GAGs was verified by selective enzyme degradation.(ABSTRACT TRUNCATED AT 400 WORDS)

Are physiological changes in capillary tube hematocrit related to alterations in capillary perfusion heterogeneity?

Average capillary tube hematocrit is hypothesized to be reduced below the mean systemic value as a result of heterogeneous distribution of blood flow among microvessels. Furthermore, capillary tube hematocrit changes with vasomotor state and these have been proposed to be due to concomitant changes in perfusion heterogeneity. We reasoned that if alterations in average capillary tube hematocrit were related to changes in perfusion heterogeneity, then mean capillary tube hematocrit should be inversely related both to the heterogeneity of capillary blood flows and to the heterogeneity of capillary hematocrit. These inferences were tested by comparing estimates of average capillary tube hematocrit (Ht) and the heterogeneities of capillary blood flow and Ht in vasoconstricted and vasodilated hamster tibialis anterior muscles. Ht was estimated from the number of red blood cells per unit capillary length (n/l); heterogeneity of capillary blood flow and heterogeneity of Ht were estimated by calculating the coefficient of variation of capillary red blood cell velocities (vrbc) and n/l, respectively. Average Ht varied with vasomotor state. However, there was no correlation between average Ht and the heterogeneity of vrbc, our index of capillary blood flow: in fact, the heterogeneity of vrbc was constant under all experimental conditions. The heterogeneity of Ht varied with vasomotor state, but not in inverse proportion to average Ht as was expected. From these observations we conclude that alterations in average capillary tube hematocrit are not due to concomitant alterations in perfusion heterogeneity.