Gene mapping by chromosome microdissection and microisolation in the chicken.

A chromosome microdissection and microisolation technique in combination with filter hybridization was developed for chromosomal localization of cloned chicken genes. The DNA was obtained from microdissected chromosome regions of metaphase spreads. Dissected DNA was amplified by polymerase chain reaction (PCR).

Association of dominant marker traits and metric traits in chickens.

This study was initiated to determine whether an allelic substitution of a dominant marker gene would identify a region close to a locus affecting expression in a metric trait. The rationale for the experiment was to utilize disequilibrium between a multiple recessive randombred Rhode Island Red (RRc) stock previously selected for quantitative trait performance and an unimproved dominant marker stock (MDM). The reporter genes in the MDM were: barring (B), silver (S), creeper (Cp), rose comb (R), double uropygial gland (U), crest (Cr), dominant white (I), frizzle (F), duplex comb (D), multiple spurs (M), polydactyly (Po), blue egg (O), pea comb (P), naked neck (Na), extended black (E), white skin (W+), muffs and beard (Mb), and feathered shanks (Fsh).

Mapping of the growth hormone gene by in situ hybridization to chicken chromosome 1.

In situ hybridization of a tritium-labeled chicken growth hormone cDNA to 62 chicken metaphase chromosome spreads was examined by analysis of silver grain distribution. A total of 554 chromosomally located grains were recorded. There was a highly significant P value for the association of silver grains to the long arm of chromosome 1.

Differential evolution in chromosomal composition during multiple cycles of subcutaneous and intravenous metastasis.

It has been proven that multiple cycles of metastasis can improve the metastatic potential and homing specificity of a tumor cell population. In the present study, verification of genetic alterations during changes in metastatic behavior was done by analyzing the chromosome composition of a methylcholanthrene induced murine fibrosarcoma, 3AM during multiple cycles of subcutaneous (SC) and intravenous (IV) metastasis. After 10 cycles of SC metastasis, a cell type, 7B, with a small t(19;19)(A;A) metacentric marker chromosome was enriched from 4% in the original population to 90% in FIOR.

Estimating meiotic disjunction frequencies in chicken translocation heterozygotes based on embryonic mortality.

Chickens heterozygous for a chromosomal translocation [MN t(1;4)] were intercrossed and the progeny were analyzed for their chromosome complement. A ratio of 1 homozygous translocation carrier to 4 heterozygous translocation carriers to 1 homozygous standard chromosome carrier was noted (n = 520), rather than the 1:2:1 ratio expected from Mendelian segregation. The excess of heterozygous carriers was apparently caused by union of complementary duplication/deficient gametes.

The improbability of irradiated spermatozoa as gene transfer vectors in chickens.

It is concluded that chromatin fragments derived from irradiated chicken spermatozoa are not viable vectors for gene transfer. In three experiments conducted at sequential intervals over a period of 1 1/2 years, no marker traits were found in 1,065 G0 progeny from irradiated spermatozoa of Minnesota Dominant Marker males inseminated into recessive Rhode Island Red and White Leghorn females. The inability to secure transformants is ascribed to the following factors: a maximum of five and probably fewer potential vector fragments for each G0 progeny because of irradiation effect on spermatozoan ability to enter the germinal disc; uncertainty of DNA integrity from highly irradiated chromatin; no known mechanism for release of chromatin fragments from irradiated spermatozoa supernumerary pronuclei; and the uncertainty of selective integration into the zygotic nucleus.

Borate G-banding technique applicable to in situ deoxyribonucleic acid/deoxyribonucleic acid hybridized chicken chromosomes.

The borate G-banding technique produced excellent longitudinal bands similar to trypsin G-bands on chicken chromosomes prepared from embryo material. Aged chromosome preparations on slides were pretreated in borate buffer (pH 9.2) for 10 to 30 s at 37 C, then stained in Giemsa solution.

Cryopreservation of semen from unique lines of chicken germ plasm.

Frozen semen is a practical means of preserving valuable germ plasm. Monitored samples of semen cryopreserved with glycerol for heterozygous, dominant marker stocks and for nine chromosomal rearrangement lines had sufficiently high fertility for germ-line retrieval. The results also indicated a potential for the genetic selection of certain lines for the freezability of spermatozoa, since stock and line differences in fertility occurred when previously frozen semen was used for insemination.

Chromosomal localization of chicken sequences homologous to the beta-actin gene by in situ hybridization.

In situ DNA/chromosome hybridization techniques were used to localize the cytoplasmic beta-actin gene in the chicken. Hybridization of a beta-actin cDNA probe to metaphase chromosome spreads indicated that sequences complementary to this probe are located on the long arm of chromosome 2 (2q) and one of chromosomes 9 through 12.

Restriction enzymes MspI, HpaII, HaeIII, and HinfI applied to chicken mitotic chromosomes.

The restriction enzymes MspI, HpaII, HaeIII, and HinfI were applied in situ to chicken metaphase chromosomes to determine if they denatured specific regions to produce banding patterns. Human metaphase chromosomes were treated simultaneously to serve as controls. The MspI, HpaII, and HaeIII enzymes produced no visible banding patterns in chicken chromosomes.

XY sex reversal syndrome in the mare: clinical and behavioral studies, H-Y phenotype.

An inherited genetic disorder causes XY embryos of the horse to develop as mares. On the basis of our study of 38 such mares, we have identified four grades or classes of XY sex reversal according to this scheme: class I, nearly normal female, of which some are fertile; class II, female with gonadal dysgenesis, normal mullerian development; class III, intersex mare with gonadal dysgenesis, abnormal mullerian development, enlarged clitoris; class IV, virilized intersex characterized by high levels of testosterone. In general, class I and class II mares were typed H-Y antigen-negative whereas class III and class IV mares were typed H-Y antigen-positive.

XY sex-reversal syndrome in the domestic horse.

The XY sex-reversal syndrome occurs when a phenotypic mare is born that has the karyotype of a stallion. The syndrome is manifested by both genotypic and phenotypic heterogeneity. The sex-reversed genetic condition occurs frequently within certain pedigrees where XY females have been found and can be readily detected by chromosome karyotyping.

Recombinant inversion chromosomes in phenotypically normal chickens.

Some progeny resulting from interbreeding of individuals heterozygous for a pericentric inversion of chromosome 1 in the chicken have the two complementary types of recombinant chromosomes arising from a single crossing-over within the inverted segment. These individuals are capable of reproduction. Their progeny can have one or the other of the two recombinant chromosomes or, if crossing-over occurs, either a normal or an inversion chromosome.

The induction and detection of diploid spermatozoa in Gallus domesticus.

Colcemid at the dose level of 0.37 mg/kg/day was injected intraperitoneally to 3 sexually active chicken males for 3 consecutive days. 10-12 days after the first colcemid injection, 14-25% of the sperm population in the semen samples from the treated males was found to be diploid in DNA content by flow microfluorometric analysis.

An autosomal recessive blind mutant in the chicken.

A type of blindness due to lack of rods and cones in the retina was found to be controlled by a single autosomal recessive gene, rc. The mutation was first identified in the second generation descendants of a male carrying an ethyl methanesulfonate (EMS) induced chromosome translocation involving one arm of the Z sex chromosome and the long (q) arm of chromosome 3. A linkage test between the locus causing the blindness and the translocation break-point on chromosome 3 was not significant.

Mapping of the genes for pea comb, blue egg, barring, silver, and blood groups A, E, H, and P in the domestic fowl.

Five chromosomal rearrangements involving chromosome 1 were utilized to test linkage relationships of the blue egg--pea comb--naked neck (O-P-Na) linkage group. An F1 stock was created by crossing rearrangement carriers with individuals carrying the traits being studied. The F1 was then backcrossed to recessive, normal chromosome tester stocks.

Induction of heteroploidy in Gallus domesticus.

Intraperitoneal colcemid injection at the dose level of 0.37 mg/kg applied 3-4 h before the first oviposition produces a high frequency of triploids among the second oviposition eggs laid the first day after colcemid treatment. The triploids, mostly 3A-ZZW type, appear to be as developmentally compatible as contemporary diploids at both embryonic and hatching level.

Sister chromatid exchange in chromosomes of cattle (Bos taurus).

Lymhocytes from clinically healthy Holstein animals (8 males and 23 females) were cultured in media containing 5-bromodeoxyuridine (10 microgram/ml) for two cycles of deoxyribonucleic acid replication (48 h). The exchange of sister chromatids per cell varied from 1 to 16 with a mean of 5.4 and a standard deviation of 2.