In vitro reactivity of digital arteries and veins to vasoconstrictive mediators in healthy horses and in horses with early laminitis

The in vitro reactivity of vasoconstrictive mediators that are implicated in acute laminitis was determined in palmar and plantar digital arteries and veins obtained from healthy horses and in palmar digital vessels of horses with early laminitis (Obel grade I). To obtain baseline reactivity data, 3 experiments were conducted, using healthy horses: (1) the reactivity of palmar and plantar digital arteries and veins to angiotensin II, norepinephrine, and 5-hydroxytryptamine (serotonin) were compared; (2) the direct effects of bacterial endotoxin on vascular reactivity were assessed; and (3) the reactivity of palmar digital arteries and veins to angiotensin II, norepinephrine, prostaglandin F2 alpha (PGF2 alpha), serotonin, and a thromboxane-endoperoxide analog (U46619) were determined. The vascular reactivity of these same 5 vasoconstrictors then was determined in horses with early laminitis and was compared with data from healthy (control) horses. Obel grade-I laminitis was experimentally induced in horses, using carbohydrate overload. Dose responses were conducted for each agent at concentrations between 10(-8)M and 10(-4)M. The potency of a drug was defined as the mean effective concentration necessary to induce 50% of maximal contraction (EC50). There were no differences in EC50 concentrations and in maximal contractions between forelimb and hind limb arteries and veins for angiotensin II, norepinephrine, and serotonin. Incubation with endotoxin had no effect on the reactivity of arteries and veins to angiotensin II, norepinephrine, and serotonin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Alterations of Endothelium-Dependent Digital Vascular Responses in Horses Given Low-Dose Endotoxin

Low doses of endotoxin cause vasoconstriction and hypoperfusion of the digit, small intestine, and cecum in horses. To determine the potential cause of these vascular alterations, in vitro vascular responses of palmar digital arteries and veins were determined in 8 horses after intravenous (IV) infusion of 1 L 0.9% NaCl (control) and 0.1 μg/kg Escherichia coli 055:B5 endotoxin in 1 L of 0.9% NaCl (endotoxin-treated). Vessels were surgically removed under general anesthesia, cut into 4-mm vascular rings, suspended in tissue baths, and attached to force displacement transducers for measurement of vascular tension. Cumulative concentration response curves to acetylcholine, bradykinin, nitroprusside, norepinephrine, 5-hydroxytryptamine (serotonin), and endothelin were determined. Maximal relaxation or contraction and the concentrations needed to produce 50% maximal relaxation or contraction were determined. Palmar digital arteries from endotoxin-treated horses relaxed significantly less in response to acetylcholine and bradykinin (endothelium-dependent), but not to nitroprusside (endothelium-independent) when compared with arteries from control horses. Digital arteries from endotoxin-treated horses also contracted significantly more with norepinephrine but less with serotonin. Digital veins responded less than digital arteries. In another study, vascular reactivity experiments documented that acetylcholine and bradykinin were endothelium-dependent vasodilators (endothelium-denuded vessels relaxed less than control vessels) in palmar digital vessels. Additionally, maximal relaxations for both vasodilators were significantly inhibited by N-nitro-L-arginine methyl ester (L-NAME), a nitric oxide antagonist, suggesting that acetylcholine and bradykinin cause relaxation through the nitric oxide pathway. The data from these studies indicate that low dose endotoxin impairs endothelium-dependent relaxation and augments adrenergic contraction of palmar digital arteries in horses.     

Palmar digital vessel relaxation in healthy horses and in horses given carbohydrate

OBJECTIVE: To compare in vitro smooth muscle relaxation of palmar digital vessels from healthy horses with those from horses in the prodromal stage of experimentally (carbohydrate) induced laminitis. ANIMALS: 16 adult horses. PROCEDURE: Segments of palmar digital vessels were obtained from 5 healthy horses and 6 horses given carbohydrate. Vascular rings from the palmar digital artery and vein were suspended in individual organ baths containing buffer solution and indomethacin; isometric tension was recorded, and contraction and relaxation were compared. Smooth muscle contraction in response to cumulative addition of phenylephrine was recorded in the absence and presence of 1 microM NG-nitro-L-arginine methyl ester (L -NAME). After wash out, vascular rings were preconstricted with phenylephrine (0.3 microM), and cumulative endothelium-dependent (acetylcholine-induced) and independent (nitroprusside-induced) smooth muscle relaxations were recorded in the absence or presence of L -NAME. RESULTS: Phenylephrine increased vascular smooth muscle tone in ring preparations of palmar digital arteries and veins. Addition of acetylcholine or nitroprusside induced relaxation of palmar digital artery and vein ring preparations. Use of L-NAME (1 microM) significantly reduced maximal relaxation induced by acetylcholine, but not by nitroprusside. Maximal relaxation induced by acetylcholine, but not by nitroprusside, was reduced in vascular rings prepared from carbohydrate-overloaded horses. CONCLUSION AND CLINICAL RELEVANCE: Reduced endothelium-dependent relaxation of palmar digital vessels may have a role in the pathophysiology of acute laminitis after carbohydrate overload in horses.     

Sonographic observations of the peripheral vasculature of the equine thoracic limb

Investigations of the equine peripheral vascular system have been constrained by the lack of a non-invasive method of examining the arteries and veins of the limbs of the conscious horse. Precise correlations were established between the gross anatomical features of the peripheral vessels and their B-mode sonographic appearance in each thoracic limb of 35 horses. A sonographic imaging protocol was established. Additional Doppler sonographic recordings defined the arterial waveforms and demonstrated that blood flow to the foot could be evaluated in the lateral proper digital artery, distal to the level of the coronary band. Valves (with 2-4 cusps) were identified in the lumina of the medial and lateral palmar common digital veins and those of the medial and lateral palmar proper digital veins. Spontaneous echo contrast, a smoke-like haze of echoic blood, was seen in the lateral and medial palmar common digital veins, the distal deep palmar venous arch and communicating branches, and the palmar proper digital veins, and occasionally seen in the distal deep palmar arterial arch and distal proper palmar digital arteries. The value of duplex sonography (B-mode and Doppler) for anatomical and physiological studies of the peripheral vasculature of the horse was clearly established. Such data could be applied to the investigation of diseases affecting the peripheral circulation.     

Effect of acute sublethal endotoxaemia on in vitro digital vascular reactivity in horses

Endotoxaemia is a syndrome linked to the development of equine laminitis; however, the relationship between them is uncertain. The aim of this experiment was to evaluate the effect of an experimental acute sublethal endotoxaemia model on in vitro equine palmar digital vascular reactivity. Rings of arteries and veins of each forelimb were obtained from 11 clinically healthy horses submitted to two surgical procedures, 3 weeks apart. Before the second surgery, 0.25 microg/kg of lipopolysaccharide from Escherichia coli O55:B5 in saline, was administered i.v. in 30 min. After 3 h, the vessels were harvested and submitted to in vitro vascular reactivity experiments and histopathology. The response to depolarizing Krebs solution (DKS, 40 mm), phenylephrine (PHE), acetylcholine (ACh) and sodium nitroprusside (SNP) were evaluated. All horses showed colic pain and watery diarrhoea, tachycardia, tachypnea, hyperthermia and leucopenia. Concentration-response curve (CRC) to PHE was shifted to the left in arteries rings from endotoxemic horses without any effect on vein rings. The CRC to ACh was shifted to the right with a reduction in the maximal response. The response to SNP and DKS was similar between groups. There was no evidence of histopathological effects. The increased response to PHE in digital arteries together with a reduction of the endothelium-dependent response to ACh in arteries and veins, confirm the existing reports where endotoxaemia was found to modify the digital vascular reactivity during the acute phase. As the digital endothelial function is impaired, there may be an increased potential to develop a digital prothrombotic state with a reduced vasodilatory capacity.     

Characterization and comparison of the responses of equine digital arteries and veins to endothelin-1

OBJECTIVE: To compare the responses of equine digital arteries (EDAs) and equine digital veins (EDVs) to endothelin-1 (ET-1) and determine the role of the endothelium and type of receptors involved in the modulation and mediation of those responses, respectively. SAMPLE POPULATION: 5 to 9 palmar digital vessels/experiment from 28 healthy horses. PROCEDURE: Rings of dissected vessels were mounted under tension between force transducer wires in organ baths containing Krebs-Henseleit solution at 30 degrees C. Responses of EDAs and EDVs (with intact [+e] or denuded [-e] endothelium) to cumulative concentrations of ET-1 (10(-10) to 3 X 10(-7) M) were compared. For (+e)EDAs and (+e)EDVs precontracted with a thromboxane-mimetic (U44069; 10(-8) M) and (-e)EDAs and (-e)EDVs, responses to an ETB receptor agonist (S6c; 10(-10) to 3 X 10(-7) M) were evaluated. Responses to ET-1 (10(-7) M) in (-e)EDAs and (-e)EDVs were evaluated after incubation with an ETA receptor antagonist (BQ-123; 3 X 10(-7) M), an ETB receptor antagonist (BQ-788; 3 X 10(-7) M), or vehicle solution. RESULTS: Endothelin-1 induced a concentration-dependent contraction of endothelium-intact and -denuded EDAs and EDVs; EDVs were more sensitive. Neither vessel type relaxed in response to S6c, although 2 of the (-e)EDAs contracted mildly. Whereas BQ-123 inhibited the (-e)EDA and (-e)EDV responses to ET-1, BQ-788 had no effect. CONCLUSIONS AND CLINICAL RELEVANCE: Endothelin-1 induced digital vasoconstriction (marked constriction in veins). This action was unaffected by endothelium and mediated predominantly by ETA receptors. These findings suggest ET-1 can induce selective digital venoconstriction.     

Evaluation of regional limb perfusion with amikacin using the saphenous,cephalic, and palmar digital veins in standing horses

Previous studies have shown that regional limb perfusion (RLP) using the palmar digital (PD) vein delivers therapeutic concentration of amikacin to the distal limb. Our hypothesis was that using the cephalic and saphenous veins for RLP will enable delivery of therapeutic concentrations of amikacin to the distal limb. Nineteen healthy horses participated in the study. The cephalic, saphenous, or PD vein was used to perfuse the limb with amikacin. Two grams of amikacin was used for RLP using the saphenous and the cephalic veins, and one gram was used in the PD vein. Synovial samples were collected from the metacarpo‐/metatarsophalangeal (MCP/MTP) joint, and blood samples were collected from the jugular vein. Maximum concentration (Cmax) of amikacin in the MCP/MTP joint using the cephalic and the saphenous vein was 277 and 363 mg/L, respectively. The amikacin concentrations achieved in the synovial fluid of the MCP/MTP joint in the current study were between 69 and 91 times the minimally inhibitory concentration of common susceptible bacterial pathogens causing orthopedic infections in horses. To conclude, this study shows that use of the proximal veins for RLP to treat distal limb infections is a viable alternative to using the palmar or plantar digital vein.     

In vitro response of large colon arterial and venous rings to vasodilating drugs in horses

OBJECTIVE: To determine in vitro vasomotor response of equine large colon arterial and venous rings with and without endothelium to vasodilator drugs, including dopamine (DOP), dopexamine (DPX), acepromazine (ACE), isoxsuprine (ISX), and nifedipine (NFP). ANIMALS: 7 adult horses. PROCEDURE: Relaxation of large colon arteries and veins in response to vasodilating drugs was determined by measuring the change in tension of vessel rings when exposed to a cumulative concentration range (10(-8) to 10(-4)M) of each drug. Vessel rings, with and without endothelium, were mounted in organ baths, attached to a transducer, and contracted with norepinephrine (NE). Cumulative concentration-response relationships, percentage maximal relaxation, and EC50 (concentration of drug required to relax the NE-induced contracted tissue to 50% of its contracted state) values were calculated. RESULTS: There were significant differences among drugs for EC50 (ACE = ISX < NFP) and percentage maximal relaxation (ACE = ISX > NFP = DPX > DOP) values in veins. Endothelium removal from veins had no significant effect. There were no differences in EC50 values for arteries; however, percentage maximal relaxation was significantly different among drugs (ACE = ISX = NFP > DPX = DOP). Endothelial removal resulted in higher EC50 and lower percentage maximal relaxation values, compared with endothelium-intact arteries. CONCLUSION AND CLINICAL RELEVANCE: ACE and ISX were the most potent and efficacious drugs evaluated and could potentially be used to improve blood flow after correction of large-colon volvulus. Dopamine cannot be recommended because of its biphasic response and potential to further decrease blood flow. Endothelium removal altered the vasodilatory responses of colonic arterial rings, but did not affect venous rings.     

Effects of Rho-kinase and Src protein tyrosine kinase inhibition on agonist-induced vasoconstriction of arteries and veins of the equine laminar dermis

OBJECTIVE: To determine the effects of inhibition of Rho-kinase or Src-family protein tyrosine kinases (srcPTK) on agonist-induced contractile responses in equine laminar arteries and veins. SAMPLE POPULATION: Laminar arteries and veins obtained from 13 adult mixed-breed horses. PROCEDURES: Laminar vessels were mounted on myographs and exposed to phenylephrine (PE), 5-hydroxytryptamine (5-HT), prostaglandin F(2) (PGF(2)), and endothelin-1 (ET-1) with or without the Rho-kinase inhibitor Y-27632 (10 microM), srcPTK inhibitor PP2 (10 microM), or a negative control analogue for PP2 (PP3; 10 microM). RESULTS: Responses to PE were reduced by use of Y-27632 in laminar vessels (approx inhibition, 55%). However, Y-27632 reduced responses to 5-HT to a greater degree in veins than in arteries (approx inhibition of 55% and 35%, respectively). The Y-27632 also reduced responses of laminar veins to ET-1 by approximately 40% but had no effect on maximum responses of laminar arteries to ET-1, although a rightward shift in the concentration response curve was evident. Addition of PP2 reduced responses to PE, 5-HT, and PGF(2) in laminar veins by approximately 40%, 60%, and 65%, respectively, compared with responses after the addition of PP3; PP2 had no effect on responses to ET-1. In laminar arteries, PP2 reduced 5-HT-induced contractions by approximately 50% but did not affect responses to PE or ET-1. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the study were consistent with activation of Rho-kinase being important during agonist-induced constriction in laminar vessels, activation of srcPTK being an agonist-dependent event, and more prominent roles for Rhokinase and srcPTK in veins than in arteries.     

Reactivity of Equine Palmar Digital Arteries and Veins to Vasodilating Agents

Palmar digital arteries and veins removed surgically from healthy horses under general anesthesia were cut into 4 mm vascular rings, suspended in tissue baths, and attached to force displacement transducers for continuous measurement of vascular tension. In vitro vascular responses were determined for acetylcholine, acepromazine, isoxsuprine hydrochloride (isoxsuprine), prostaglandin E₂ (PGE₂), and prostaglandin I₂ (prostacyclin). After preconstriction with norepinephrine hydrochloride (norepinephrine), or prostaglandin F₂ alpha (PGF₂ alpha), the concentrations needed to produce 50% maximum relaxation (EC₅₀) and the maximum percentage of relaxation were determined for each drug.
Acetylcholine was the most potent arterial vasodilator (smallest EC₅₀ value) and PGE₂ was the least potent. Prostacyclin was the least potent venodilator (highest EC₅₀ value); there were no differences between acetylcholine, acepromazine, isoxsuprine, and PGE₂. Isoxsuprine produced greater arterial relaxation than all other agents. Isoxsuprine and acepromazine produced significantly greater venous relaxation than did acetylcholine and PGE₂. Prostacyclin produced minimal vasodilation of arteries or veins. Acepromazine and isoxsuprine relaxed the veins significantly more than the arteries. When PGF₂ alpha was used instead of norepinephrine to preconstrict the arteries and veins, the potency and effectiveness of acepromazine and isoxsuprine to produce vasodilation were significantly decreased. Results indicate that acepromazine and isoxsuprine can relax the equine digital vasculature but their effectiveness varies depending on the origin of the constriction.     

Antagonism in isolated equine digital vessels of contraction induced by epinephrine in the presence of hydrocortisone and an aqueous extract of black walnut (Juglans nigra)

Prazosin, isoxsuprine, and nifedipine were screened for ability to reverse contraction of isolated equine digital vascular strips produced by epinephrine (Epi) in the presence of hydrocortisone (Hc) and an aqueous extract of black walnut (Juglans nigra) (BW). Two arteries and two veins from each of three horses for each drug (n = 9) were maintained in isolated tissue baths in Krebs' bicarbonate buffer with 95% oxygen at 37 degrees C. Six-point Epi concentration-response (C-R) curves were obtained for each vessel in the presence of Hc, BW, and the appropriate vehicle. This was repeated for each vessel using one of two concentrations of one of the three test drugs. Each drug and concentration combination was tested on a total of three arteries and three veins. Prazosin produced a concentration-dependent shift of the Epi C-R curve to the right but the curve maintained the same maximum height and slope, which is consistent with competitive alpha 1 adrenergic blockade. Isoxsuprine exhibited similar behavior, although the precise mechanism of action for isoxsuprine is unknown. Conversely, nifedipine did not shift the curve but did depress maximum contraction, suggesting a non-competitive interaction consistent with its mechanism of calcium-channel blockade.     

Endoscopy of the Digital Flexor Tendon Sheath in Horses

An arthroscopic procedure for examination of the digital flexor tendons and tendon sheath was developed in 16 equine limbs and 12 horses. Distension of the tendon sheath and insertion of the arthroscope was accomplished through a cul-de-sac on the palmar or plantar surface of the tendon sheath 1 to 2 cm palmar or plantar to the digital neurovascular structures and between the annular ligament and proximal digital annular ligament. A single arthroscope entry point allowed examination of all regions of the tendon sheath cavity and most surfaces of the digital flexor tendons within the sheath. Distal to the fetlock, surgical procedures could be performed through additional entry portals on the lateral, medial, or palmar surfaces of the tendon sheath. The palmar digital vessels and nerves were avoided by palmar placement of the instrument incisions and insertion of a needle before incising the skin. The fetlock canal and proximal regions of the tendon sheath were examined by redirecting the arthroscope. Flexion of the fetlock aided passage of the arthroscope into the proximal tendon sheath regions. Evaluation of the palmar surface of the superficial digital flexor tendon was limited by the midline attachment of the tendon sheath, otherwise the surfaces of the tendons and tendon sheath could be examined with 25 degrees and 70 degrees arthroscopes. The tendon sheath was more tightly invested to the tendons in the proximal regions, limiting the arthroscope movements and second instrument access.     

Use of computed tomography angiography for the evaluation of a cutaneous haemangioma in a Standardbred horse

Cutaneous haeangiomas are benign vascular neoplasms that arise from endothelial cells of blood vessels. Haemangiomas account for 0.6%–4% of all equine cutaneous neoplasms and the fetlock is the most commonly affected site. We describe the use of computed tomography angiography (CTA) for the evaluation of a cutaneous haemangioma located on the plantarolateral aspect of the left hindlimb fetlock of a 9-month-old Standardbred colt. Computed tomography angiography of the affected fetlock was performed under general anaesthesia. The medial plantar artery was catheterised and a total volume of 50 ml of iodinated non-ionic contrast medium (Iopamidol, 300 mg I/ml, Bracco Imaging Canada) was injected at a rate of 2 ml/s. Following contrast medium administration, the dorsal metatarsal artery and branches including the lateral and medial digital arteries were well demarcated. Two smaller lateral and medial arteries were also identified, forming the vascular network of the metatarsophalangeal joint. At the level of the haemangioma, two tortuous arteries arising from the lateral digital artery were identified, in addition to multiple small branches from nearby cutaneous arteries. These vessels supplied the homogeneously strongly contrast-enhancing cutaneous mass. The initial goal of the CTA study was to map the vascular anatomy for arterial embolisation in conjunction with pharmacological therapy. Considering the involvement of multiple small arterial branches, complete surgical excision along with ligation of the two main supplying arteries was alternatively elected, resulting in a successful long-term outcome.     

Effect of voltage-gated and capacitative calcium entry blockade on agonist-induced constriction of equine laminar blood vessels

OBJECTIVE: To characterize the relative contributions of voltage-gated and capacitative Ca(2+) entry to agonist-induced contractions of equine laminar arteries and veins. ANIMALS: 16 adult mixed-breed horses. PROCEDURES: Laminar arteries and veins were isolated and mounted on small vessel myographs for the measurement of isometric tension. Concentration-response curves were obtained for the vasoconstrictor agonists phenylephrine, 5-hydroxytryptamine (5-HT), prostaglandin F(2) (PGF(2)), and endothelin-1 (ET-1) either in the absence of extracellular Ca(2+) or in the presence of the voltage-gated Ca(2+) channel inhibitor diltiazem or the putative inhibitor of capacitative Ca(2+) entry, trifluoromethylphenylimidazole. RESULTS: In the absence of extracellular Ca(2+), maximal responses of veins to 5-HT, phenylephrine, ET-1 and PGF(2) were reduced by 80%, 50%, 50%, and 45%, respectively; responses of arteries to 5-HT, phenylephrine, and ET-1 were reduced by 95%, 90%, and 20%, respectively. Although diltiazem did not affect the maximal responses of veins to any agonist, responses of arteries to 5-HT, phenylephrine, and ET-1 were reduced by 40%, 50%, and 27%, respectively. Trifluoromethylphenylimidazole did not affect maximal responses of veins, but did reduce their contractile responses to low concentrations of ET-1 and PGF(2). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the contribution of extracellular Ca(2+) to laminar vessel contractile responses differs between arteries and veins and also between contractile agonists, voltage-gated Ca(2+) entry is more predominant in laminar arteries than in veins, and capacitative Ca(2+) entry has a minor role in agonist-induced contractile responses of laminar veins.     

Tenoscopic examination and proximal annular ligament desmotomy for treatment of equine "complex" digital sheath tenosynovitis

OBJECTIVE: To determine the outcome of horses with "complex" digital tenosynovitis treated by tenoscopic proximal annular ligament desmotomy and resection of synovial masses or adhesions, or both, within the digital sheath. STUDY DESIGN: Retrospective evaluation. ANIMALS OR SAMPLE POPULATION: Twenty-five horses with a clinical and ultrasonographic diagnosis of palmar or plantar proximal annular ligament constriction and ultrasonographic evidence of synovial masses or adhesions within the digital tendon sheath. METHODS: Each horse had tenoscopic surgery for annular ligament desmotomy combined with adhesiolysis and/or synovial mass resection. Mean follow-up time was 3.4 years. Spearman's rank correlation was used to assess the relationship between functional outcome or cosmetic results and preoperative variables including duration of clinical signs, digital sheath synovial fluid total protein concentration and nucleated cell count, thickness of the palmar or plantar proximal annular ligament (PAL), severity of adhesions, severity of synovial masses, degree of synovial distention, or limb affected. RESULTS: A total of 18 (72%) horses returned to athletic soundness, 4 were improved but not sound, and 3 were not improved. Cosmetic outcome was normal in 10 horses, improved but not normal in 12, and not improved in 3 horses. Cosmetic and functional outcome were significantly adversely affected by the duration of clinical signs and the severity of synovial masses. CONCLUSIONS: With appropriate tenoscopic surgical attention, horses with complex tenosynovitis syndrome characterized by synovial masses, adhesions, or both adhesions and masses, and PAL constriction, have a good prognosis for return to athletic soundness. CLINICAL RELEVANCE: Horses with PAL constriction and additional digital tendon sheath pathology such as adhesions and synovial masses have a 72% chance of returning to sound athletic performance, however 60% of horses retain some degree of cosmetic blemish in the affected limb. There is an inverse relationship between the duration of clinical signs and outcome, and therefore, prompt surgical attention is advised.     

Evaluation of activation of protein kinase C during agonist-induced constriction of veins isolated from the laminar dermis of horses

OBJECTIVE: To determine the effects of the protein kinase C (PKC) inhibitor, Ro-31-8220, on agonist-induced constriction of laminar arteries and veins obtained from horses. SAMPLE POPULATION: Laminar arteries and veins obtained from 8 adult mixed-breed horses. PROCEDURES: Laminar arteries and veins were isolated and mounted on small vessel myographs for the measurement of isometric tension. Concentration-response curves were then obtained for the vasoconstrictor agonists phenylephrine, 5-hydroxytryptamine, prostaglandin F(2), and endothelin-1. All responses were measured with or without the addition of Ro-31-8220 (3 microM). RESULTS: Laminar veins were more sensitive to vasoconstrictor agonists than laminar arteries, and incubation of laminar veins with Ro-31-8220 resulted in significantly smaller agonist-induced contractile responses for all agonists tested. In contrast, Ro-31-8220 had no effect on agonist-induced contractile responses of laminar arteries. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the study were consistent with activation of PKC being confined to agonist-induced contraction of laminar veins isolated from the laminar dermis of horses. Consequently, the possible involvement of PKC in the venoconstriction observed during the development of laminitis is worthy of further investigation.     

Effects of induction of capacitative calcium entry on equine laminar microvessels

OBJECTIVE: To determine the effects of induction of capacitative Ca2+ entry on tone in equine laminar arteries and veins. SAMPLE POPULATION: Laminar arteries and veins from 6 adult mixed-breed horses. PROCEDURE: Arteries and veins were isolated and mounted on small vessel myographs for the measurement of isometric tension. Capacitative Ca2+ entry was induced by incubating the vessels with the specific Ca2+-ATPase inhibitor thapsigargin (100nM) in a Ca2+-free physiologic salt solution. Capacitative Ca2+ entry-associated contractile responses were determined by the subsequent addition of 2mM Ca2+ to the solution bathing the vessels; in some experiments, either the voltage-gated Ca2+ blocker diltiazem (10microM) or the putative capacitative Ca2+ entry inhibitor trifluoromethylphenylimidazole (300microM) was added to the bathing solution 15 minutes prior to a second 2mM Ca2+ exposure. The Sr2+ permeability of the capacitative Ca2+ entry pathway in laminar vessels was assessed by exposing the vessels to 4mM Sr2+ after induction of capacitative Ca2+ entry with thapsigargin. RESULTS: Induction of capacitative Ca2+ entry elicited robust contractile responses in laminar veins but did not increase tone in laminar arteries. In laminar veins, capacitative Ca2+ entry-induced contractile responses were unaffected by preincubation with diltiazem, attenuated by trifluoromethylphenylimidazole, and were impermeable to Sr+. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that induction of capacitative Ca2+ entry elicits vasoconstriction in equine laminar veins but not in laminar arteries and should therefore be considered a potential mechanism by which selective venoconstriction occurs in horses during the development of acute laminitis.