Black hair follicular dysplasia, an autosomal recessive condition in dogs.

Using histology, a coat color abnormality and the subsequent hair loss were diagnosed as black hair follicular dysplasia. A pedigree analysis of an affected litter and literature review suggests that this is inherited as an autosomal recessive trait. The melanocyte stimulating hormone receptor gene is ruled out by using linkage analysis.

In situ hybridization of five loci to cattle chromosome 1.

The genes for pituitary-specific transcription factor (PIT1), propionyl coenzyme A carboxylase, beta-polypeptide (PCCB), transferrin (TF), trichohyalin (THH), and involucrin (IVL) were mapped to cattle chromosome 1 (BTA 1) by isotopic in situ hybridization. Two of the loci were mapped from cattle PCR products and three from human ATCC probes. PIT1 localized to segment 1q2; PCCB to 1q3; and TF, THH, and IVL to 1q4. These localizations agree with the homology previously shown between BTA 1 and human chromosome 3 (HSA 3). Some homology with HSA 1 has been established with the mapping of THH and IVL to BTA 1q4.

Fertility effects of the 14;20 Robertsonian translocation in cattle.

Superovulation and embryo collection procedures were used to study the effect of the 14;20 Robertsonian translocation on fertility and embryo viability. Karyotypes were successfully completed on cells from 77 of the 279 embryos prepared for such analysis. Embryos from 4 cows heterozygous for the translocation were studied. Two bulls with the same condition were studied by using their semen in artificial insemination of cows with normal karyotypes. The proportions of fertilized ova and transferable embryos were not different between cows with the 14;20 translocation and those with normal karyotypes, indicating that fertilization rates were not affected by the translocation. Twenty-two percent of the embryos which were karyotyped had an unbalanced karyotype and would theoretically not have survived to term. All of the theoretically predicted chromosome complements from such a translocation were observed as were three 58,XX,t karyotypes and a 58,XX karyotype. There was no difference in the percentage of embryos with abnormal karyotypes whether the cow or bull was the carrier. Results therefore indicate that fertility is rather severely impaired in carriers of the 14;20 translocation, as was observed with the 1;29 translocation, with most loss due to embryo mortality rather than a lowered conception rate.

Chromosomal aneuploidy associated with spontaneous abortions and neonatal losses in cattle.

Pericardial sac samples from 77 bovine aborted fetuses and stillborn calves were submitted for tissue culture; cells from 55 of these samples were grown successfully in culture. Six of the 55 karyotyped fetuses (10.7%) had an abnormal chromosome complement, in 3 of which (5.5%) the abnormality was probably the cause of death. This level of abnormality is relatively high when one considers that most fetuses were >8 months gestational age. Approximately 5-7% of human stillbirths and 50% of first-trimester aborted fetuses have chromosome anomalies. If a similar situation exists in cattle, as suggested by these data, chromosome abnormalities may be a major cause of early fetal loss in cattle. Most chromosomally abnormal fetuses had multiple malformations, which suggests that the diagnostic use of chromosome analysis is most cost effective for malformed fetuses and newborns. Twins were present in a higher proportion of these fetuses than expected based on their incidence among liveborn cattle.

Physical mapping of SOD1 to bovine chromosome 1.

Superoxide dismutase 1 (SOD1) was mapped to cattle chromosome 1q12 --> q14 by in situ methods. Both traditional in situ hybridization using tritium and a new technique, direct in-situ single copy PCR (DISC-PCR), were used in two separate laboratories. Both human and bovine SOD1 clones were tritium labeled for radioactive in situ hybridization. A primer pair based on the bovine SOD1 gene (Barendse et al., 1994b) was used for the DISC-PCR procedure. The map location of SOD1 is close to collagen 6A1. SOD1 is a potentially important type 1 anchor locus in the region where the gene for horns in cattle was recently mapped (Georges et al., 1993; Schmutz et al., 1995).

DNA marker-assisted selection of the polled condition in Charolais cattle.

Five Charolais families known to segregate for both horned and polled were selected and tested for linkage analysis by use of microsatellites and karyotyping for Robertsonian translocation 1;29. No recombinants were found between any of these markers and the polled phenotype or each other. When statistical analysis was performed, the logarithm of the odds (LOD) indicated that there was 100% linkage occurring between the markers and the phenotype (p < 0.001). These microsatellite markers, TGLA49 and BM6438, can be assumed to be very close to the actual gene that determines the polled phenotype. Another linked marker, SOD1, was physically mapped, which places all of these markers within 1q12-14, very near the centromere of Chromosome (Chr) 1. A homozygous polled cow was identified in this study by following the alleles at both markers and the phenotypes in her family.

Gene mapping from a bovine 1;29 DNA library prepared with chromosome microdissection.

Bovine gene mapping is progressing rapidly using syntenic group mapping based on somatic cell hybrids and linkage, and to a lesser extent on in situ hybridization. Single chromosome DNA libraries are a logical next step, and this was, therefore, the aim of our laboratory. Since we have access to several cattle with t(1;29) and this chromosome is readily distinguishable, we chose this as our first target--recognizing that we would not produce a "single" chromosome library in the strict sense because two autosomes are represented. We utilized an inverted microscope and a micromanipulator fitted with glass instruments pulled specifically to dissect off approximately 100 t(1;29) chromosomes per microdrop. A glass chamber made to accommodate a hanging drop was used to extract the DNA under a dissecting microscope. The DNA was then cleaved with EcoRI and inserted in lambda gtwes arms. Host cells were then infected with these phage and positive clones obtained. The first clone, isolated from this library by hybridization with a human collagen 6A1 cDNA, was mapped by in situ hybridization to bovine Chromosome some (Chr) 1q12-q14, near the centromere. The second clone, an anonymous DNA fragment (D1S11), was mapped to 1q43-q46, near the terminal end.

Detection of leucochimaerism in bovine twins by DNA fingerprinting.

Karyotyping and hypervariable genetic markers indicate extensive leucochimaerism between pairs of dizygotic twins in cattle, a result of placental vascular anastomosis. The extent of this chimaerism includes both kind and number of cells exchanged. All heterosexual twin pairs harboured two types of leucocytes, having either XX or XY chromosome pairs, and 30 of 31 pairs of twins shared identical DNA fingerprints. Although chromosome results from skin fibroblasts indicate that some chimaerism occurs in the skin, the low level allows for differentiation of genotypes between twins. The results warrant against the common practice of using blood samples for DNA typing if twinning is not properly documented.

Embryonic loss in superovulated cattle caused by the 1;29 Robertsonian translocation.

The effect of the 1;29 Robertsonian translocation on fertility was studied using embryos resulting from matings of nine carrier cows and two carrier bulls. Embryos were collected from the following three mating groups utilizing superovulation: normal bull cross normal cow, normal bull cross translocation carrier cow, and translocation carrier bull cross normal cow. The proportion of ova which were fertilized did not vary among the groups, indicating that fertilization rates were not affected by the translocation. The translocation cows did yield fewer embryos on average than did cows with normal karyotypes, which may suggest ovulation rates are reduced (at least after superovulation attempts) in cattle carrying the 1;29 translocation. Twenty of 39 embryos successfully karyotyped had abnormal chromosome complements. All four of the theoretically predicted karyotypes and two additional abnormal combinations were found. Eight of 39 (20.5%) embryos karyotyped had unbalanced karyotypes which would have resulted in embryonic loss. The proportion of embryos with unbalanced karyotypes, was slightly higher when the cow (36%) carried the translocation than when the bull (19%) did. Results of this study indicate that fertility is impaired due to the presence of this translocation. The major loss in reproductive potential appears to be due to embryonic loss rather than fertilization failure.