Preventing experimental vertical transmission of scrapie by embryo transfer.

This study investigated whether the transmission of naturally occurring scrapie in sheep can be prevented using embryo transfer. Embryos were collected from 38 donor ewes in a Suffolk sheep flock with a high incidence of naturally occurring scrapie, treated with a sanitary procedure (embryo washing) recommended by the International Embryo Transfer Society and then transferred to 58 scrapie-free recipient ewes. Ninety-four offspring were produced. None of the offspring or the recipient ewes developed scrapie. Furthermore, offspring derived from embryos collected from donor ewes bred to the immunohistochemically positive ram did not develop scrapie. We conclude that scrapie was not transmitted to offspring via the embryo nor was the infective agent transmitted to recipient ewes during embryo transfer procedures.

The association of MHC genes with autism.

Several immune abnormalities have been noted in autistic subjects. These associations have been extended to the Major Histocompatibility Complex (MHC), a section of DNA remarkable for the number of encoded proteins with immunological functions. The strongest MHC association identified thus far is for the null allele of C4B in the class III region. The complex allelic composition of C4 as determined by immunoelectrophoresis is discussed. Low levels of C4 resulting from the null allele may be important in disease pathogenesis especially since C4 has been identified in developing brain neurons. The DNA region just telomeric to C4 has several genes including tumor necrosis factor which encode proteins with immunological functions. These proteins may act in concert with C4 in disease contribution and the genes should be more closely examined.

Localization of the locus causing Spider Lamb Syndrome to the distal end of ovine Chromosome 6.

Spider Lamb Syndrome (SLS) is a semi-lethal congenital disorder, causing severe skeletal abnormalities in sheep. The syndrome has now been disseminated into several sheep breeds in the United States, Canada, and Australia. The mode of inheritance for SLS is autosomal recessive, making the identification and culling of carrier animals difficult due to their normal phenotype. Two large pedigrees segregating for the SLS mutation were established, and a genome scan with genetic markers from previously published genome maps of cattle and sheep was used to map the locus causing SLS. Genetic linkage between SLS and several microsatellite markers, OarJMP8, McM214, OarJMP12, and BL1038, was detected, thereby mapping the SLS locus to the telomeric end of ovine Chromosome (Chr) 6. Alignment of ovine Chr 6 with its evolutionary ortholog, human Chr 4, revealed a positional candidate gene, fibroblast growth factor receptor 3 (FGFR3).

Association of genes within the major histocompatibility complex with attention deficit hyperactivity disorder.

The objective was to determine whether a relationship exists among the complement C4B gene, a DR region gene and attention deficit hyperactivity disorder (ADHD). Thirty-one subjects with ADHD, their mothers, all but 5 of their fathers, and 90 normal subjects living in northern Utah were studied. DR and C4B typing were performed by serologic HLA typing techniques and the DNA methods PCR-RFLP. The alleles of 2 genes, the null allele of the C4B gene and the beta 1 allele of the DR gene, encode for products involved in immune function and regulation. Each of these alleles was found to be significantly associated with ADHD. Moreover, approximately 55% of the ADHD subjects carried both of these alleles on 1 of their chromosomes, compared to only 8% of normal controls. Genes related to the immune system may be associated with development of the symptoms of ADHD.

Strong association of the third hypervariable region of HLA-DR beta 1 with autism.

We reported that the major histocompatibility complex (MHC) including the null allele of the C4B gene and the extended haplotype B44-C30-DR4 is associated with autism. We report now that the third hypervariable region (HVR-3) of certain DR beta 1 alleles have very strong association with autism. The HVR-3 of DR beta 1* 0401 or the shared HVR-3 alleles DR beta 1* 0404 and DR beta 1* 0404 and DR *0101, was expressed on extended haplotypes in 23 of 50 (46%) autistic subjects as compared to only 6 of 79 (7.5%) normal subjects. Another HVR-3 sequence, the DR beta 1* 0701 allele, was carried on extended haplotypes in 16 (32.0%) of the autistic subjects as compared to 8 (10.1%) of the normal subjects.

Immunogenetic studies in autism and related disorders.

The major histocompatibility complex comprises a number of genes that control the function and regulation of the immune system. One of these genes, the C4B gene, encodes a product that is involved in eliminating pathogens such as viruses and bacteria from the body. We previously reported that a deficient form of the C4B gene, termed the C4B null allele (no C4B protein produced) had an increased frequently in autism. In this study we attempted to confirm the increased incidence of the C4B null allele in autism and investigated the presence of a C4B null allele in two other childhood disorders, attention-deficit hyperactivity disorder and dyslexia (reading disability). In addition, we explored the relationship of autism to the DR beta 1 gene, a gene located close to the C4B in autism. We confirmed the finding of an increased frequency of the C4B null allele in autism and found that the related disorders also had an increased frequency of this null allele. In addition, two alleles of the DR beta 1 gene also had significantly increased representation in the autistic subjects.

Is decreased blood plasma concentration of the complement C4B protein associated with attention-deficit hyperactivity disorder?

OBJECTIVE: The complement system is a group of blood proteins that play an important role in defending against viral and bacterial infections. The objective of this investigation was to study the plasma levels of the C4B protein in attention-deficit hyperactivity disorder (ADHD) in an attempt to associate infections with the development of some cases of this disorder. METHOD: C4B plasma protein levels were studied using an enzyme-linked immunosorbent assay in a group of 23 subjects meeting DSM-III-R criteria for ADHD and a similar number of age- and sex-matched controls. Also studied were parents of the ADHD subjects. RESULTS: C4B plasma levels (157.0 micrograms/mL) in the ADHD subjects were significantly (p < .01) lower than those (239.3 micrograms/mL) in the normal age-matched subjects. Mothers of the ADHD subjects also had significantly lower C4B values compared with mothers of normal children. On the other hand, C4B values in the fathers were not significantly altered. CONCLUSIONS: Decreased C4B levels in ADHD, if replicated, may represent an important marker for ADHD (or a subgroup of ADHD). It also seems plausible that C4B levels are an important etiological factor for ADHD.

Increased frequency of the extended or ancestral haplotype B44-SC30-DR4 in autism.

Autism likely results from several different etiologies or a combination of pathological mechanisms. Recent studies suggest that this disorder may be associated with immune abnormalities, pathogen-autoimmune processes and perhaps the major histocompatibility complex (MHC). In a preliminary study we found that 22 autistic subjects had an increased frequency of the extended or ancestral MHC haplotype B44-SC30-DR4. The current study attempted to confirm this observation by studying 23 additional randomly chosen autistic subjects, most of their parents and 64 unrelated normal subjects. In agreement with earlier findings B44-SC30-DR4 was associated with autism. In combining the data from the original and current studies, B44-SC30-DR4 or a substantial fragment of this extended haplotype was represented in 40% of the autistic subjects and/or their mothers as compared to about 2% of the unrelated subjects. It is concluded that one or more genes of the MHC is (are) involved in the development of some cases of autism.

Characterization of a strain of cerebral endothelial cells derived from goat brain which retain their differentiated traits after long-term passage.

A strain of cerebral endothelial cells was established from isolated cortical microvessels of caprine brain. These cells, which are referred to as EC1 cells, can be routinely subcultured to 32 passages without the loss of differentiated morphologic and immunologic traits. The ability to routinely subculture EC1 cells is an important asset, given that isolated cerebral endothelial cells in mammals generally lose their differentiated traits after only 2 to 3 passages. EC1 cells were shown to contain Factor VIII-related antigen, which is a specific marker for cells of endothelial origin. EC1 cells morphologically demonstrated a scarcity of pinocytotic vesicles on their apical surfaces, a lack of trans-cytoplasmic vesicles, and the ability to form in culture confluent monolayers with tight junctional complexes. Therefore, EC1 cells possess specific antigenic and ultrastructural features which classify them as being small vessel endothelial cells of the blood-brain barrier type. Cytogenetic evaluation of EC1 cells demonstrated a normal female goat 60,XX karyotype and confirmed the apparent non-transformed nature of EC1 cells due to the lack of chromosome abnormalities or rearrangements. Using scanning electron microscopy, EC1 cells were also shown to form confluent monolayers on mixed nitrocellulose filters, a feature that will enable the development of an in vitro system to study trans-endothelial transport. Given that EC1 cells are readily subcultured and grow well on nitrocellulose filters, and that they resemble cerebral endothelium in vivo, it seems evident that EC1 cells can be used as a versatile model for the study of blood-brain barrier function, regulation, and pathology.

Prevention of scrapie transmission in sheep, using embryo transfer.

Reciprocal embryo transfers were made between scrapie-inoculated and scrapie-free sheep (Cheviot and Suffolk breeds) to measure scrapie transmission via the embryo (using offspring from embryos of scrapie-inoculated donors and scrapie-free recipients) and via the uterus (using offspring from embryos of scrapie-free donors and scrapie-inoculated recipients taken by cesarean section). Two control groups of offspring, 1 from scrapie-free parents (negative) and 1 from scrapie-inoculated parents (positive), also were included. All sheep were observed for clinical signs of scrapie until death or for a minimum of 60 months. Final diagnosis was made on the basis of histopathologic findings or results of mouse inoculation and/or proteinase-K-resistant protein analysis. Thirty to 61% of the scrapie-inoculated donor/recipient sheep within groups developed scrapie within 8 to 44 months after inoculation. None of the scrapie-free donor/recipients, including those gestating embryos from scrapie-inoculated donors, developed scrapie. Also, none of the offspring observed to > or = 24 months of age from reciprocal cross, via embryo (0/67), or via the uterus (0/25), or from the negative-control group (0/33) developed scrapie. Fifty-six of the offspring via embryo, 19 of these via the uterus, and 31 negative controls survived to > or = 60 months of age. Of the 21 sheep in the positive-control group, 2 (9.5%) developed scrapie, 1 at 31 months of age and 1 at 42 months of age. In the Cheviot offspring, the percentage of sheep carrying the short incubation allele ranged from 24 to 44% and the percentage in the Suffolk offspring ranged from 61 to 83%.(ABSTRACT TRUNCATED AT 250 WORDS)

Polymorphisms of a scrapie-associated fibril protein (PrP) gene and their association with susceptibility to experimentally induced scrapie in Cheviot sheep in the United States.

The duration of the incubation period for scrapie, a fatal transmissible neurodegenerative disorder of sheep and goats, is mainly determined by the Sip gene, which has 2 alleles (sA--susceptible and pA--resistant). A diagnostic test is not available to detect scrapie in live animals. We analyzed genomic DNA extracted from frozen sheep brains collected from Cheviot sheep of the United States that had been inoculated with the SSBP/1 scrapie inoculum. Digestion of the DNA with EcoRI or HindIII followed by the addition of a scrapie-associated fibril protein (PrP)-specific marker probe, yielded fragments of 6.8 (e1) and 4.0 (e3) kb, or 5.0 (h1) and 3.4 (h2) kb, respectively. Fragments e1 and h2 were associated with the histopathologic diagnosis of scrapie, and fragments e3 and h1 were associated with survival. A valine/alanine polymorphism within the PrP coding region that resulted in a BspHI site was further used to determine the genotype of these Cheviot sheep. Digestion of polymerase chain reaction fragments with BspHI resulted in an undigested fragment b- (0.840 kb), digested fragments b+ (0.460 and 0.380 kb), or both types of fragments. Survival time of b+/b+ homozygous sheep was significantly (P < 0.01) shorter (218 +/- 26.0 days) than survival time for b-/b- sheep (> 700 days after inoculation). Results indicated that b+ and b- are markers for the Sip sA and pA alleles, respectively. The intermediate duration of the incubation period for heterozygous sheep (b+/b-; 342.9 +/- 25.3 days) indicated that the Sip sA allele is expressed codominantly to the Sip pA allele.

True hermaphroditism in a wild sheep: A clinical report.

Intersexuality in sheep is rare, with the freemartin anomaly being the most common. We describe here a true hermaphrodite in a wild sheep. An F(1) wild sheep ewe of Argali-mouflon X Mexican desert bighorn breeding was bred to an F(1) ram of the same breeding. A single lamb was born with the external appearance of a normal female. The lamb grew faster than its female cohorts, and by 6 months of age exhibited the aggressive behavior, size, coloration and horn development associated with males. Phenotypically, the intersex had female external genitalia with an enlarged clitoris. A human chorionic gonadotrophin (hCG) response test was performed when the intersex was 1-year-old and serum testosterone, progesterone and estradiol levels were compared to the response of a normal female and male of similar age and breeding. An exploratory celiotomy revealed two gonadal-like structures associated with a female reproductive tract. Histopathology of the structures revealed spermatogenically inactive testicular vessels and ovarian tissue with primary follicles. The reproductive tract was complete with two uterine horns and a cervix. The intersexuality is attributed to an XX/XXY mosaic.

High-resolution G-banded karyotype and idiogram of the goat: a sheep-goat G-banded comparison.

An elongated G-banded karyotype and idiogram of the domestic goat (Capra hircus) is presented. Sheep (Ovis aries)-goat comparison suggests the terminal light band (Xq 2 9) present in the goat is absent in the sheep.