Disorders of sexual development and associated changes in the pituitary-gonadal axis in dogs.

Normal sexual differentiation depends on completion of chromosomal sex determination, gonadal differentiation, and development of the phenotypic sex. An irregularity in any of these three steps can lead to a disorder in sexual development (DSD). We examined nine dogs with DSD by abdominal ultrasonography, laparotomy, histologic examination of the gonads, and reproductive tract, cytogenetic analysis, and mRNA expression of the SRY gene. We also determined the plasma concentrations of luteinizing hormone (LH), estradiol-17ß, and testosterone before and after administration of gonadotropin-releasing hormone (GnRH) and compared these results with those obtained in anestrous bitches and male control dogs. The gonads of three dogs with DSD contained both testicular and ovarian tissue, while in the other six only testicular tissue was found. Each of the dogs had a uterus. Based on gynecologic examination, cytogenetic analysis, and the histology of the gonads, seven of the nine dogs appeared to be XX sex reversals. Three of these were XX true hermaphrodites and four were XX males; the other two dogs had incomplete XY gonadal dysgenesis. All seven XX sex-reversed dogs were found to be negative for the SRY gene by polymerase chain reaction. The basal plasma luteinizing hormone (LH) concentration was significantly higher in dogs with DSD than in anestrous bitches but not significantly different from that in male dogs. The basal plasma LH concentration increased significantly after GnRH administration in all dogs with DSD. The basal plasma estradiol concentration was significantly higher in dogs with DSD than in anestrous bitches but not significantly different from that in male dogs. The basal plasma testosterone concentration was lower in dogs with DSD than in male dogs. In all dogs with DSD both the basal and GnRH-induced plasma testosterone concentrations were above the upper limit of their respective ranges in the anestrous bitches. In conclusion, the secretion of LH and estradiol in these dogs with DSD, all of which had testicular tissue in their gonads, was similar to that in male control dogs. These results indicate that the basal and/or GnRH-stimulated plasma testosterone concentration might be used to detect the presence of testicular tissue in dogs with DSD.

Persistent Mullerian duct syndrome in a Miniature Schnauzer dog with signs of feminization and a Sertoli cell tumour.

A 5-year-old male Miniature Schnauzer was presented with unilateral cryptorchidism and signs of feminization. Abdominal ultrasonography revealed an enlarged right testis and a large, fluid-filled cavity that appeared to arise from the prostate. Computed tomography revealed the cavity to be consistent with an enlarged uterine body, arising from the prostate, and showed two structures resembling uterine horns that terminated close to the adjacent testes. The dog had a normal male karyotype, 78 XY. Gonadohysterectomy was performed and both the surgical and the histological findings confirmed the presence of a uterus in this male animal, resulting in a diagnosis of persistent Mullerian duct syndrome (PMDS). The enlarged intra-abdominal testis contained a Sertoli cell tumour. Computed tomography proved to be an excellent diagnostic tool for PMDS.

Minimal external masculinization in a SRY-negative XX male Podenco dog.

Normal mammalian sex differentiation takes place in three genetically controlled steps: chromosomal sex determination (XX or XY), gonadal differentiation and development of the phenotypic sex. Animals are considered to be sex reversed if chromosomal sex determination and gonadal development are not in agreement. In this report, sex reversal is described in a 1.5-year-old Podenco dog that was referred because of suspected recurrent growth of a previously removed os clitoridis in the vulva. With that exception the dog was phenotypically female, but had never been in oestrus and exhibited male behaviour. Abdominal ultrasonography showed a small tubular structure dorsal to the bladder, consistent with a uterus. An ovoid structure resembling a gonad was visible between the right kidney and inguinal canal. Plasma testosterone concentrations before and after GnRH administration indicated the presence of functional testicular tissue. Two testes, each with its epididymis and ductus deferens, and a complete bicornuate uterus were removed surgically. Cytogenetic analysis of peripheral blood lymphocytes showed a normal female karyotype (78, XX). These findings are consistent with the diagnosis of an XX male. PCR analysis of genomic DNA revealed that the SRY gene was absent. In summary, this report describes the first SRY-negative XX male Podenco dog with an almost complete female phenotype despite high basal and stimulated plasma testosterone concentrations. It is hypothesized that the clinical observations in this dog may have been caused by reduced and delayed Müllerian-inhibiting substance secretion and the absence of conversion of testosterone to dihydrotestosterone due to 5alpha-reductase deficiency.

Intravitreal triamcinolone for the treatment of refractory macular edema in idiopathic intermediate or posterior uveitis.

PURPOSE: Cystoid macular edema (CME) is the most significant cause of visual loss associated with idiopathic uveitis. The authors report on the use of intravitreal triamcinolone acetonide (IVTA) in a group of patients with macular edema due to idiopathic intermediate and posterior uveitis. METHODS: Retrospective, noncomparative, interventional case series. Thirty-three eyes were included with uveitic CME that was refractory to topical steroids, oral prednisone, or a combination thereof. Previous steroid treatment did not result in elevated intraocular pressure (IOP). The eyes received an intravitreal injection with 10 mg triamcinolone acetonide, after best-corrected visual acuity (BCVA) and fluorescein angiography (FA) were assessed. Ophthalmologic examination including FA was regularly performed during a 1-year follow-up period. RESULTS: Within 12 weeks after injection of IVTA, 50% of the eyes responded with an improvement in vision of more than two lines and 30% of the eyes reached an IOP of > or = 21 mmHg (p<0.01). All eyes with an elevated IOP responded well on topical antiglaucoma medication. After 12 months follow-up 40% of the eyes responded with an improvement in vision of more than two lines and 28% of the affected eyes underwent phacoemulsification during the follow-up. No other complications occurred within a year after the treatment. CONCLUSIONS: In macular edema due to idiopathic intermediate or posterior uveitis IVTA improves the visual acuity within the first 3 months. However, thereafter the visual acuity decreases again. Cataract and elevated IOP are common side effects.

A radiation hybrid map of the X-chromosome of the dog (Canis familiaris).

The dog serves as an animal model for several human diseases including X-chromosome diseases. Although the canine X-chromosome is one of the largest chromosomes in the dog, only a few markers have been mapped to it to date. Using a commercially available canine whole genome radiation hybrid (RH) panel we have localized 14 microsatellite markers, 18 genes and 13 STSs on the canine X-chromosome, extending the total number of mapped markers to 45 covering an estimated 830 cR. Out of these 45 markers, seven distinct groups of markers could be established with an average spacing of 18.8 cR(3000) and ten markers remained unlinked. Using FISH analysis, six markers could be mapped physically to the p- or q-arm of the X-chromosome. Combined with the FISH mapping, three RH groups could be assigned to the p-arm and two RH groups to the q-arm. Comparison with the human X-chromosome map revealed conserved synteny up to 234 cR (TIMP1-ALAS2-AR-IL2RG-XIST). We show here that the similarity of the canine and human X-chromosomes is the largest for any mammalian species beyond the primates.

Mapping of the Na+, K(+)-ATPase subunit alpha 2 (ATP1A2) and muscle phosphofructokinase (PFKM) genes in pig by somatic cell hybrid analysis.

Two expressed sequence tags were isolated from a porcine skeletal muscle cDNA library and identified as the putative partial cDNAs of the porcine Na+, K(+)-ATPase subunit alpha 2 (ATP1A2) and muscle phosphofructokinase (PFKM) genes after sequencing and homology search. Results of analysis of a pig-rodent somatic cell hybrid panel by PCR allowed the assignments of ATP1A2 to porcine chromosome (chr) 4 and of PFKM to porcine chr 5. These assignments support previously observed conservation of syntenic relationships between human chr 1 and porcine chr 4 and between human chr 12 and porcine chr 5.

Analysis of chromosome aberrations in a mammary carcinoma cell line from a dog by using canine painting probes.

A cell line derived from a spleen metastasis of a mammary carcinoma in a female dog was analyzed by fluorescence in situ hybridization with canine chromosome-specific paints. The cell line showed a modal chromosome number of 77, with three (90% of the cells) or four (10% of the cells) biarmed chromosomes. Aberrations observed relate to chromosomes 8 or 11, 13 or 15, 37, 38, and X and include chromosome loss (X), formation of isochromosomes (8 or 11, 13 or 15), and centric fusion (37 and 38). In all aberrations, whole chromosomes are involved. None of the genes known to be related to breast cancer development in humans and that has mapped in the dog is located on one of the aberrant chromosomes. The results of this study show that chromosome painting is a most useful tool for the analysis of canine tumor cells.

Reproduction of the beet cyst nematode Heterodera schachtii Schm. on transformed root cultures of Beta vulgaris L.

Transformed hairy root cultures of Beta vulgaris L., grown in petri-dishes, were inoculated with a suspension of surface-sterilized larvae of the beet cyst nematode Heterodera schachtii Schm. Larvae developed into both male and female adults. Juvenile larvae were hatched from the newly-formed cysts, indicating that fertilization had occurred. Results of a glasshouse test showed that H. schachtii did not lose its pathogenicity after being cultured on transformed roots. This technique can be developed further for the mass-propagation of sedentary nematodes and for the in vitro storage of isclates.