Analysis In Dogs Research Articles

Serum cortisol concentration and force plate analysis in the assessment of pain associated with sodium urate–induced acute synovitis in dogs

To determine the relationship between serum cortisol concentration and pain severity as measured by force platform gait analysis in dogs with experimentally induced synovitis of the stifle joint. 10 healthy hound-type dogs. Dogs underwent 2 study phases. In the first phase, serum cortisol concentration, systolic arterial blood pressure, heart rate, and gait data were obtained at 0 (first sample), 2.5, 5, 7.5, and 10 hours. In the second phase, the same data were gathered immediately before (0 hours) and 2.5, 5, 7.5, and 10 hours after induction of acute urate synovitis in the left stifle joint. Data were statistically evaluated to compare changes in variable values over time and to determine the accuracy of serum cortisol measurements for diagnosis of acute orthopedic pain. Following induction of synovitis, ground reaction forces were significantly decreased relative to preinduction values at 2.5, 5.0, 7.5, and 10.0 hours and serum cortisol concentration was significantly increased at 2.5 hours. A cortisol concentration of >or= 1.6 microg/dL indicated pain with a 91% sensitivity and 35% specificity. In this model, cortisol concentration may be useful for diagnosing pain in dogs. Although, with a cutoff of >or= 1.6 microg/dL, pain would be detected in most dogs with pain, some pain-free dogs would also be identified as having pain. Conversely, dogs with a serum cortisol of < 1.6 microg/dL would be unlikely to have pain. Validation of this diagnostic test in a large, heterogeneous group of clinical patients is necessary.

Basal measures of insulin sensitivity and insulin secretion and simplified glucose tolerance tests in dogs

There is need for simple, inexpensive measures of glucose tolerance, insulin sensitivity, and insulin secretion in dogs. The aim of this study was to estimate the closeness of correlation between fasting and dynamic measures of insulin sensitivity and insulin secretion, the precision of fasting measures, and the agreement between results of standard and simplified glucose tolerance tests in dogs. A retrospective descriptive study using 6 naturally occurring obese and 6 lean dogs was conducted. Data from frequently sampled intravenous glucose tolerance tests (FSIGTTs) in 6 obese and 6 lean client-owned dogs were used to calculate HOMA, QUICKI, fasting glucose and insulin concentrations. Fasting measures of insulin sensitivity and secretion were compared with MINMOD analysis of FSIGTTs using Pearson correlation coefficients, and they were evaluated for precision by the discriminant ratio. Simplified sampling protocols were compared with standard FSIGTTs using Lin's concordance correlation coefficients, limits of agreement, and Pearson correlation coefficients. All fasting measures except fasting plasma glucose concentration were moderately correlated with MINMOD-estimated insulin sensitivity (| r| = 0.62–0.80; P < 0.03), and those that combined fasting insulin and glucose were moderately closely correlated with MINMOD-estimated insulin secretion ( r = 0.60–0.79; P < 0.04). HOMA calculated using the nonlinear formulae had the closest estimated correlation ( r = 0.77 and 0.74) and the best discrimination for insulin sensitivity and insulin secretion (discriminant ratio 4.4 and 3.4, respectively). Simplified sampling protocols with half as many samples collected over 3 h had close agreement with the full sampling protocol. Fasting measures and simplified intravenous glucose tolerance tests reflect insulin sensitivity and insulin secretion derived from frequently sampled glucose tolerance tests with MINMOD analysis in dogs.

Comparison of Biomaterial Implants in the Dental Socket: Histological Analysis in Dogs

Bone graft procedures have been used commonly in buco-maxillo-facial surgery. For this reason, many researchers have evaluated the bone substitutes. The present study evaluated soft and hard tissue reactions to two different hydroxyapatites HAs (synthetic HA and natural HA) and bioactive glass implanted into the sockets immediately after extraction. First and third upper and lower premolars, on both sides, were extracted from six female dogs. The alveolar sockets were randomly assigned to four groups: Group 1 - control (unfilled), Group 2 - filled with synthetic hydroxyapatite, Group 3 - filled with bovine bone mineral (natural HA), and Group 4 - filled with bioactive glass. The animals were euthanized at 4 weeks (n = 2), 8 weeks (n = 2), and 28 weeks (n = 2) after extraction. The mandible and maxilla of each animal were removed for histological analysis to determine soft tissue reactions, newly formed bone, bone characteristics, and presence or absence of implanted materials. Most particles of synthetic hydroxyapatite had bone formation on their surface, although some particles showed a layer of fibrous connective tissue. The bovine bone mineral group exhibited particles partially replaced with bone formation. The bioactive glass group showed particles with a thin layer of calcified tissue, but was absent in some specimens, suggesting complete resorption. All biomaterials had similar behavior. Bovine bone mineral, compared to synthetic hydroxyapatite and bioactive glass, showed a larger number of particles covered with osseous tissue. All biomaterials interfered with the socket repair process.

Application of Gel Permeation HPLC for Lipoprotein Profiling in Dogs

A high performance liquid chromatography system with a gel permeation column (GP-HPLC) and an on-line dual enzymatic system was applied to lipoprotein analysis in dogs. A high density lipoprotein (HDL) fraction obtained by conventional ultracentrifugation gave a single peak at around 28-29 min. Similarly, a low density lipoprotein (LDL) fraction gave single peak at around 24-25 min. The lipoprotein profiles of healthy dogs were contained large HDL peaks and small LDL peaks, and VLDL and CM were only marginally detected. In diabetic dogs, concentrations of VLDL-triglyceride and VLDL-total-cholesterol were elevated significantly. The lipoprotein profile analysis by GP-HPLC method would be useful in explication of abnormality of lipid metabolism in dogs.

Influence of implant positioning in extraction sockets on osseointegration: histomorphometric analyses in dogs

To evaluate the influence of implant positioning into extraction sockets on osseointegration. Implants were installed immediately into extraction sockets in the mandibles of six Labrador dogs. In the control sites, the implants were positioned in the center of the alveolus, while in the test sites, the implants were positioned 0.8 mm deeper and more lingually. After 4 months of healing, the resorptive patterns of the alveolar crest were evaluated histomorphometrically. All implants were integrated in mineralized bone, mainly composed of mature lamellar bone. The alveolar crest underwent resorption at the control as well as at the test sites. After 4 months of healing, at the buccal aspects of the control and test sites, the location of the implant rough/smooth limit to the alveolar crest was 2 + or - 0.9 mm and 0.6 + or - 0.9 mm, respectively (P<0.05). At the lingual aspect, the bony crest was located 0.4 mm apically and 0.2 mm coronally to the implant rough/smooth limit at the control and test sites, respectively (NS). From a clinical point of view, implants installed into extraction sockets should be positioned approximately 1 mm deeper than the level of the buccal alveolar crest and in a lingual position in relation to the center of the alveolus in order to reduce or eliminate the exposure above the alveolar crest of the endosseous (rough) portion of the implant.

Tooth Replantation After Use of Euro-Collins Solution or Bovine Milk as Storage Medium: A Histomorphometric Analysis in Dogs

Euro-Collins solution was developed for the preservation of organs for transplantation, whose characteristics have raised interest for its use as a storage medium for avulsed teeth before replantation. This study evaluated histologically and morphometrically the healing process of dog teeth replanted after storage in Euro-Collins solution or bovine milk. Eighty roots of 4 young adult mongrel dogs were randomly assigned to 4 groups (n = 20) and the root canals were instrumented and obturated with gutta-percha and a calcium hydroxide-based sealer. After 2 weeks, the teeth were extracted and subjected to the following protocols: GI (negative control), replantation immediately after extraction; GII (positive control), bench-drying for 2 hours before replantation; GIII and GIV, immersion in 10 mL of whole bovine milk and Euro-Collins solution at 4 degrees C, respectively, for 8 hours before replantation. The animals were sacrificed 90 days postoperatively. The pieces containing the replanted teeth were subjected to routine processing for histologic and histometric analyses under light microscopy and polarized light microscopy. Root resorption was observed in all groups. GII exhibited the greatest loss of dental structure (P < .01), and inflammatory resorption was predominant in this group. Storage in milk showed poorer results than immediate replantation and storage in Euro-Collins solution (P < .01). The teeth stored in Euro-Collins solution presented similar extension of root resorption and periodontal ligament reorganization to those of immediately replanted teeth. The findings of this study suggest that the Euro-Collins solution is an adequate storage medium for keeping avulsed teeth for up to 8 hours before replantation.

Immunophenotyping and Gene Rearrangement Analysis in Dogs with Lymphoproliferative Disorders Characterized by Small-Cell Lymphocytosis

Lymphocytosis caused by neoplastic proliferation of small lymphocytes is occasionally difficult to distinguish by morphological examination from nonneoplastic lymphocytosis. To examine the clinical utility of gene rearrangement analysis for demonstrating neoplastic proliferation of small lymphocytes, gene rearrangement analysis was performed in comparison with immunophenotyping using peripheral lymphocytes in dogs with small lymphocytosis. Thirty-one dogs with small-cell lymphocytosis (8,100-884,300/microl) were enrolled. By immunophenotyping, lymphocytosis of all dogs was suggested to be neoplastic in nature based on the detection of marked expansion of phenotypically homogeneous lymphocytes or the presence of an aberrant antigen-expressing population of lymphocytes. In contrast, gene rearrangement analysis represented clonality in 27 dogs (detection rate of 87%). From the present study, gene rearrangement analysis was considered to be worthwhile to strengthen the evidence of neoplastic proliferation of small lymphocytes when coupled with immunophenotyping and to be a suitable diagnostic substitute if immunophenotyping is not available in clinical practice.

Bone repair in mandibular body osteotomy after using 2.0 miniplate system – histological and histometric analysis in dogs

The objective of this study was to evaluate the bone repair along a mandibular body osteotomy after using a 2.0 miniplate system. Nine adult mongrel dogs were subjected to unilateral continuous defect through an osteotomy between the mandibular 3rd and 4th premolars. Two four-hole miniplates were placed in accordance with the Arbeitgeimeinschaft für Osteosynthesefragen Manual. Miniplates adapted to the alveolar processes were fixed monocortically with 6.0-mm-length titanium alloy self-tapping screws, whereas miniplates placed near the mandible bases were fixed bicortically. At 2, 6 and 12 weeks, three dogs were sacrificed per period, and the osteotomy sites were removed, divided into three thirds (Tension Third, TT; Intermediary Third, IT; Compression Third, CT) and prepared for conventional and polarized light microscopy. At 6 weeks, while the CT repaired faster and showed bone union by woven bone formation, the TT and IT exhibited a ligament-like fibrous connective tissue inserted in, and connecting, newly formed woven bone overlying the parent lamellar bone edges. At 12 weeks, bone repair took place at all thirds. Histometrically, proportions of newly formed bone did not alter at TT, IT and CT, whereas significantly enhanced bone formation was observed for the 12-week group, irrespective of the third. The results demonstrated that although the method used to stabilize the mandibular osteotomy allowed bone repair to occur, differences in the dynamics of bone healing may take place along the osteotomy site, depending on the action of tension and compression forces generated by masticatory muscles.

Fatores relacionados com a positividade de cães para leishmaniose visceral em área endêmica do Estado do Rio Grande do Norte, Brasil

O presente estudo objetivou descrever alguns fatores relacionados à positividade de cães para leishmaniose visceral, em área endêmica do Estado do Rio Grande do Norte. Foram analisados 198 cães, dos quais 62 foram recrutados de propriedades rurais e 136 de residências urbanas. O diagnóstico foi realizado através dos testes de imunofluorescência indireta e imunoadsorção enzimática. Dos cães analisados, 45% dos positivos eram provenientes da área rural e 35% da área urbana. Analisando a função dos animais, a prevalência foi de 50% dos cães de guarda nas áreas rural e de 43% na área urbana. Quanto aos cães que tinham contato com outros cães de origem rural ou urbana, a prevalência observada de foi 54% e 32%, respectivamente. No que se refere ao sexo, as fêmeas da zona rural foram mais prevalentes. Estes resultados sugerem uma atenção maior sobre o papel destes animais como reservatório dessa zoonose, como também na urbanização da leishmaniose visceral e o inquérito epidemiológico desta doença, no qual estas informações poderão contribuir para o planejamento de estratégias de controle do cão como reservatório doméstico.

Arthrocentesis and synovial fluid analysis in dogs and cats

ARTHROCENTESIS is the aspiration of synovial fluid from a joint space for diagnostic evaluation. The technique is simple to perform for most peripheral appendicular joints in small animals and is...

Biochemical Analysis of Pericardial Fluid and Whole Blood in Dogs with Pericardial Effusion

Studies evaluating pericardial fluid analysis in dogs to determine the etiology of pericardial effusions have yielded conflicting results. The purpose of this prospective study was to compare acid-base status, electrolyte concentrations, glucose, and lactate of pericardial fluid to peripheral blood from dogs with pericardial effusion and to compare these variables between dogs with neoplastic and nonneoplastic pericardial effusion. Acid-base status, electrolyte concentrations, glucose, hematocrit, urea nitrogen, and lactate concentrations were evaluated in peripheral blood samples and in pericardial effusion samples of 41 client-owned dogs with pericardial effusion. Common abnormal findings in the peripheral blood of dogs with pericardial effusion included hyperlactatemia (n = 38 [of 41]; 93%), hyponatremia (n = 25/41; 61%), hyperglycemia (n = 13/41; 32%), and hypermagnesemia (n = 13/41; 32%). Bicarbonate, sodium, ionized calcium, glucose, and hematocrit were all significantly lower in the pericardial fluid compared with peripheral blood, whereas lactate, chloride, and PCO2 were significantly higher in the pericardial fluid. When comparing the concentrations of variables in the pericardial fluid of dogs with neoplasia (n = 28) to those without neoplasia (n = 13), pH, bicarbonate, and chloride were significantly lower in dogs with neoplasia, whereas lactate, hematocrit, and urea nitrogen were significantly higher in the pericardial fluid of dogs with neoplasia. The difference between peripheral and pericardial glucose concentrations was significantly larger in dogs with neoplasia than in dogs without neoplasia. Although differences between variables in dogs with neoplastic and nonneoplastic pericardial effusion were documented, clinical relevance is likely limited by the degree of overlap between the 2 groups.

Regenerative Potential of Platelet‐Rich Plasma Added to Xenogenic Bone Grafts in Peri‐Implant Defects: A Histomorphometric Analysis in Dogs

The purpose of this investigation was to evaluate the regenerative influence of platelet-rich plasma (PRP) added to xenogenic bone grafts on bone histomorphometric parameters in a dog model. Ninety endosseous dental implants were inserted in the mandibles of nine hound dogs. Mesial and distal 3-wall peri-implant defects were surgically created adjacent to the implants. Defects were randomly assigned to three groups: demineralized freeze dried bone with platelet-rich plasma (DFDB + PRP), DFDB alone, and no treatment (NT). Animals were sacrificed at 1, 2, and 3 months according to a previously established randomization schedule, and specimens were subjected to histomorphometric analysis. Percentages of bone area inside the implant threads (BiIT), bone-to-implant contact (BIC), and bone area outside implant threads (BoIT) were recorded. Treatment effects were evaluated using analysis of variance models. The effect of the three treatments on the outcome measures did not differ significantly by healing time (P > 0.05 for the healing time by treatment interaction). However, the average (standard deviation) percentage of BIC and BoIT was significantly different between the treatment groups. In particular, the average percentage of BIC differed between peri-implant defects treated with DFDB + PRP (33.8% [11.0]) and those treated with DFDB alone (28.5% [10.8]; P = 0.042), as well as those in the NT group (27.9% [11.0]; P = 0.024). Furthermore, the average percentage of BoIT differed significantly between defects treated with DFDB + PRP compared to defects in the NT group (51.6% [12.2] versus 43.3% [10.3]; P = 0.005). There was borderline evidence to suggest that the average percentage of BiIT and BIC was significantly different depending on the length of the healing time (P = 0.054 and P = 0.085, respectively). This study found that the addition of PRP to xenogenic bone grafts demonstrated a low regenerative potential in this animal model.

Biochemical Analysis of Pericardial Fluid and Whole Blood in Dogs with Pericardial Effusion

Studies evaluating pericardial fluid analysis in dogs to determine the etiology of pericardial effusions have yielded conflicting results. The purpose of this prospective study was to compare acid-base status, electrolyte concentrations, glucose, and lactate of pericardial fluid to peripheral blood from dogs with pericardial effusion and to compare these variables between dogs with neoplastic and nonneoplastic pericardial effusion. Acid-base status, electrolyte concentrations, glucose, hematocrit, urea nitrogen, and lactate concentrations were evaluated in peripheral blood samples and in pericardial effusion samples of 41 client-owned dogs with pericardial effusion. Common abnormal findings in the peripheral blood of dogs with pericardial effusion included hyperlactatemia (n = 38 [of 41]; 93%), hyponatremia (n = 25/41; 61%), hyperglycemia (n = 13/41; 32%), and hypermagnesemia (n = 13/41; 32%). Bicarbonate, sodium, ionized calcium, glucose, and hematocrit were all significantly lower in the pericardial fluid compared with peripheral blood, whereas lactate, chloride, and PCO2 were significantly higher in the pericardial fluid. When comparing the concentrations of variables in the pericardial fluid of dogs with neoplasia (n = 28) to those without neoplasia (n = 13), pH, bicarbonate, and chloride were significantly lower in dogs with neoplasia, whereas lactate, hematocrit, and urea nitrogen were significantly higher in the pericardial fluid of dogs with neoplasia. The difference between peripheral and pericardial glucose concentrations was significantly larger in dogs with neoplasia than in dogs without neoplasia. Although differences between variables in dogs with neoplastic and nonneoplastic pericardial effusion were documented, clinical relevance is likely limited by the degree of overlap between the 2 groups.

Clinical Applicability and Limitations of Peripheral Venous Blood samples for blood gas analysis in Dogs

Clinical Applicability and Limitations of Peripheral Venous Blood samples for blood gas analysis in Dogs

Evaluation of postoperative respiratory function by serial blood gas analysis in dogs treated with transdermal fentanyl

AbstractObjective: To perform blood gas analysis of the respiratory response to transdermal fentanyl in dogs which have experienced an open‐chest surgical procedure.Design: Prospective trial.Setting: Veterinary Teaching Hospital Surgical Research and Student Laboratory.Intervention: Dogs were purchased for a surgical laboratory. Students performed a cranial abdominal exploratory and diaphragmatic hernia repair. Sixteen dogs were divided into 2 groups. Dogs received transdermal fentanyl (group F), using an average dose of 4.8 µg/kg/hr, applied to the caudal‐lateral abdomen 22 hours before surgery, or intravenous buprenorphine (group B; 0.02 mg/kg) given 1 hour prior to anesthetic induction and every 6 hours postoperatively. All dogs received intravenous acepromazine (0.05 mg/kg) preoperatively and every 6 hours postoperatively. Dogs were instrumented with carotid artery catheters.Measurements and main results: Arterial blood gas values were analyzed every 2 hours postoperatively. Plasma fentanyl levels were analyzed every 4 hours postoperatively. The mean carbon dioxide tension (PCO2) did not exceed 45 mmHg in either group. The range in mean PCO2 levels was 32.9 (± 3.4)−38.1 (± 3.9) in group B and 34.7 (± 3.25)−43.6 (± 5.5) in group F. At 2 time points, the mean PCO2 was significantly lower in group B compared with normal levels in group F. Hypoxemia occurred in both the groups. The range in mean oxygen tension (PO2) was 76.5 (± 18.3)−91.1 (± 16.3) in group B and 76.0 (± 10.8)−96.6 (± 7.6) in group F. There was no significant difference in PO2 between groups. Levels of fentanyl considered to be analgesic were maintained for the postoperative period.Conclusions: The use of a relatively high dose of transdermal fentanyl did not induce postoperative hypoventilation as evidenced by serial arterial blood gas analysis in this model.

Effects of carprofen on renal function and results of serum biochemical and hematologic analyses in anesthetized dogs that had low blood pressure during anesthesia.

To investigate effects of IV administered carprofen on indices of renal function and results of serum biochemical and hematologic analyses in dogs anesthetized with acepromazine-thiopentone-isoflurane that had low blood pressure during anesthesia. 6 healthy Beagles. A randomized crossover study was conducted, using the following treatments: saline (0.9% NaCl solution)-saline, saline-carprofen, and carprofen-saline. Saline (0.08 ml/kg) and carprofen (4 mg/kg) were administered IV. The first treatment was administered 30 minutes before induction of anesthesia and immediately before administration of acepromazine (0.1 mg/kg, IM). Anesthesia was induced with thiopentone (25 mg/ml, IV) and maintained with inspired isoflurane (2% in oxygen). The second treatment was administered 30 minutes after onset of inhalation anesthesia. Blood gases, circulation, and ventilation were monitored. Renal function was assessed by glomerular filtration rate (GFR), using scintigraphy, serum biochemical analyses, and urinalysis. Hematologic analysis was performed. Statistical analysis was conducted, using ANOVA or Friedman ANOVA. Values did not differ significantly among the 3 treatments. For all treatments, sedation and anesthesia caused changes in results of serum biochemical and hematologic analyses, a decrease in mean arterial blood pressure to 65 mm Hg, an increase of 115 pmol/L in angiotensin II concentration, and an increase of 100 seconds in time required to reach maximum activity counts during scintigraphy. Carprofen administered IV before or during anesthesia did not cause detectable significant adverse effects on renal function or results of serum biochemical and hematologic analyses in healthy Beagles with low blood pressure during anesthesia.

Effects of different variables on whole blood cholinesterase analysis in dogs.

The influence of several variables such as sample and reagent storage, anticoagulants, reaction temperature, pH, and substrate concentration on whole blood cholinesterase determination was studied. Storage of nondiluted whole blood samples at room temperature or under refrigeration (4 C) was adequate for short-term storage (3 days to 2 weeks). However, freezing would be more appropriate for long-term storage (> or = 1 month), and successive thawing and freezing did not produce any loss of cholinesterase activity. All reagents (2,2'-dithiodipyridine as chromophore and acetylthiocholine and butyrylthiocholine as substrates) were stable for 3 months when frozen. Heparin and ethylenediaminetetraacetic acid were the most suitable anticoagulants for whole blood acetylcholinesterase and butyrylcholinesterase determination, because citrate yielded lower acetylcholinesterase values and fluoride inhibited butyrylcholinesterase. Increases in reaction temperature and pH yielded higher cholinesterase values but also increased nonenzymatic substrate hydrolysis. Higher cholinesterase and nonenzymatic substrate hydrolysis values were obtained as higher substrate concentrations were used.

Effects of platelet clumping on platelet concentrations measured by use of impedance or buffy coat analysis in dogs.

To determine whether platelet clumps are homogeneously distributed in blood samples, and whether platelet concentrations (PC) obtained by use of impedance and buffy coat analysis can be considered minimum values when platelet clumps are present. Prospective study. 50 blood samples obtained from 30 dogs. 10 blood samples containing platelet clumps were used and 10 smears were made from each sample; amount of platelet clumping was graded for all 100 smears. Blood from each of 20 healthy dogs was divided between 2 EDTA tubes before and after platelet clumping was induced by adenosine diphosphate (ADP). The PC for each ADP-treated and untreated sample were measured, using impedance and quantitative buffy coat analyzers. Platelet clumps were evident in all 100 blood smears, but the amount of clumping varied considerably within some samples. Using the impedance analyzer, the PC of ADP-treated samples were significantly lower and never higher than the PC of untreated samples. Using the buffy coat analyzer, some ADP-treated samples had increased PC; however, significant differences were not detected between treated and untreated samples. Platelet clumping was not homogeneous within blood samples. When platelet clumps were identified by direct examination of blood smears, the PC detected by use of the impedance analyzer could be considered minimum values. In contrast, the PC detected by use of the buffy coat analyzer were sometimes increased. Useful information can be obtained by measuring PC in blood with platelet clumps; values obtained by use of impedance can be considered minimums, and values obtained by use of buffy coat analysis may be either minimum values or reasonable estimates of PC.

Canine TCOF1; cloning, chromosome assignment and genetic analysis in dogs with different head types.

We describe the construction of a dog embryonic head/neck cDNA library and the isolation of the dog homolog of the Treacher Collins Syndrome gene, TCOF1. The protein shows a similar three-domain structure to that described for human TCOF1, but the dog gene lacks exon 10 and contains two exons not present in the human sequence. In addition, exon 19 is differentially spliced in the dog. How these structural differences relate to TCOF1 phosphorylation is discussed. Isolation of a genomic clone allowed the exon/intron boundaries to be characterized and the dog TCOF1 gene to be mapped to CF Chr 4q31, a region syntenic to human Chr 5. Genetic analysis of DNA of dogs from 13 different breeds identified nine DNA sequence variants, three of which gave rise to amino acid substitutions. Grouping dogs according to head type showed that a C396T variant, leading to a Pro117Ser substitution, is associated with skull/face shape in our dog panel. The numbers are small, but the association between the T allele and brachycephaly, broad skull/short face, was highly significant (p = 0.000024). The short period of time during which the domestic dog breeds have been established suggests that this mutation has arisen only once in the history of dog domestication.

The canine homeobox gene MSX2: sequence, chromosome assignment and genetic analysis in dogs of different breeds.

The MSX2 gene encodes a homeodomain transcription factor important for normal head and face morphogenesis. MSX2 is expressed in key craniofacial structures during development and mutations in the human gene give rise to various craniofacial abnormalities. We are interested in the genetic basis of non-pathogenic variation in skull and face shape. As part of this study we have analysed DNA from a panel of different dog breeds, selected for the differences they show in these traits and investigated MSX2 as a candidate gene. In this paper we describe the cloning of the canine homologue of MSX2, the determination of its structure, sequence and localization of the gene to dog chromosome 4q23. The DNAs from 11 individual domestic dogs belonging to 10 different breeds were sequenced in a search for genetic variation. Our studies show that variation in MSX2 does not contribute to the diversity of face shape observed in these domestic dogs and that the MSX2 sequence is strongly conserved between different dog breeds. The proximal promoter shows a high level of interspecies sequence conservation and several conserved transcription factor binding motifs have been identified and their significance discussed.