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X-linked congenital stationary night blindness (CSNB) is a recessive non-progressive retinal disorder characterized by night blindness, decreased visual acuity, myopia, nystagmus and strabismus. Two distinct clinical entities of X-linked CSNB have been proposed. Patients with complete CSNB show moderate to severe myopia, undetectable rod function and a normal cone response, whereas patients with incomplete CSNB show moderate myopia to hyperopia and subnormal but measurable rod and cone function. The electrophysiological and psychophysical features of these clinical entities suggest a defect in retinal neurotransmission. The apparent clinical heterogeneity in X-linked CSNB reflects the recently described genetic heterogeneity in which the locus for complete CSNB (CSNB1) was mapped to Xp11.4, and the locus for incomplete CSNB (CSNB2) was refined within Xp11.23 (ref. 5). A novel retina-specific gene mapping to the CSNB2 minimal region was characterized and found to have similarity to voltage-gated L-type calcium channel alpha1-subunit genes. Mutation analysis of this new alpha1-subunit gene, CACNA1F, in 20 families with incomplete CSNB revealed six different mutations that are all predicted to cause premature protein truncation. These findings establish that loss-of-function mutations in CACNA1F cause incomplete CSNB, making this disorder an example of a human channelopathy of the retina.

Background: Mutations in CACNA1F, which encodes the Cav1.4 subunit of a voltage-gated L-type calcium channel, cause X-linked incomplete congenital stationary night blindness (CSNB2), a condition of defective retinal neurotransmission which results in night blindness, reduced visual acuity, and diminished ERG b-wave. We have characterized two putative murine CSNB2 models: an engineered null-mutant, with a stop codon (G305X); and a spontaneous mutant with an ETn insertion in intron 2 of Cacna1f (nob2).

Tuberous sclerosis (TSC) is a multisystem autosomal dominant hamartosis whose genetics is complicated by reduced penetrance and widely varying clinical expression. Results of linkage analyses have variously suggested two different locations for a TSC gene. A collaborative dataset has been assembled to clarify the issue of genetic heterogeneity. We have now analyzed the data from a combined sample of 111 families. Using Ott's HOMOG programs, we completed three tests of homogeneity: (1) for chromosome 9q, (2) for chromosome 11q, and (3) for the combined 9q and 11q data. For test 1 the chi-square (1 df) was 21.54 (p less than 0.001), for test 2 the chi-square (1 df) was 0.13 (p greater than 0.35), and for test 3 the chi-square (2 df) was 37.61 (p less than 0.0001). Additionally, we examined the combined data for evidence that a third, as yet unlinked locus exists. Results of this last test were suggestive but not significant. Clearly loci for TSC are present on both chromosomes 9q and 11q. The maximum likelihood estimate of the proportion of chromosome 9q-linked families is 0.38, for chromosome 11q-linked families is 0.47, and for the unlinked type 0.15. Alternative explanations for these latter families include chance sampling of recombinants, nongenetic phenocopies, or misclassification.

Incomplete X-linked congenital stationary night blindness (CSNB) is a recessive, non-progressive eye disorder characterized by abnormal electroretinogram and psychophysical testing and can include impaired night vision, decreased visual acuity, myopia, nystagmus, and strabismus. Including the 20 families previously reported (Bech-Hansen et al. 1998b), we have now analyzed patients from a total of 36 families with incomplete CSNB and identified 20 different mutations in the calcium channel gene CACNA1F. Three of the mutations account for incomplete CSNB in two or more families, and a founder effect is clearly demonstrable for one of these mutations. Of the 20 mutations identified, 14 (70%) are predicted to cause premature protein truncation and six (30%) to cause amino acid substitutions or deletions at conserved positions in the alpha1F protein. In characterizing transcripts of CACNA1F we have identified several splice variants and defined a prototypical sequence based on the location of mutations in splice variants and comparison with the mouse orthologue, Cacnalf.

Congenital stationary night blindness (CSNB) is a nonprogressive retinal disorder characterized by night blindness, nystagmus, myopia, a variable decrease in visual acuity, an abnormal electroretinographic response, and a disturbance in dark adaptation. Two forms of X-linked CSNB have been defined, complete CSNB in which rod function is extinguished, and incomplete CSNB in which rod function is reduced but not extinguished, as seen by electroretinography and dark adaptometry. In studying a large family of Mennonite ancestry, we have confirmed linkage between the locus (CSNB2) for incomplete CSNB and genetic markers in the Xp11 region. In particular, lod scores of 12.25 and 15.26 at zero recombination were observed between CSNB2 and the markers DXS573 and DXS255. Detailed analysis of critical recombinant chromosomes in this extended family have refined the minimal region for the CSNB2 locus to the interval between DXS6849 and DXS8023 in Xp11.23.

The identification by fundus examination of those females carrying an X-linked retinitis pigmentosa (RP) gene can reportedly be as high as 87%. In genetic counselling sessions with young females with a 50% risk of being a carrier who wished to know their status, it has not been possible to achieve such a level of success. A review and reanalysis of previous reports indicated that if a tapetal-like reflex was not present in those age 35 years or less, the likelihood of identifying a carrier by fundus examination was small. A family with 7 females with a 50% risk of being a carrier of X-linked RP was evaluated using haplotype analysis in an attempt to identify the X chromosome carrying the RP gene. In the family described, it was possible to establish that a mutation in the RP3 locus most likely causes the disease. This has permitted the determination of the carrier status in each of the females with a high degree of certainty.

Linkage between X-linked congenital stationary night blindness (CSNB1) and seven markers on the X chromosome was investigated in a large four-generation Albertan kindred. We detected significant linkage between the CSNB1 locus and the locus DXS255 (maximum lod score = 6.73 at a recombination fraction of 6%; confidence interval of 1% to 18%), which anchors the CSNB1 locus to the proximal region near p11.22 on the short arm of the X chromosome.

Four nuclear and two cytoplasmic chloramphenicol-resistance markers were selected in S. cerevisiae. The effect of the suppressive factor (abnormal mitochondria deoxyribonucleic acid) on the inheritance of these markers was studied. Nuclear markers were shown to be unaffected by the suppressive factor, in contrast to the loss of cytoplasmically inherited chloramphenicol resistance upon the generation of the suppressive factor.

Genetic analyses of 48-hr-old zygote-daughter-colony cells from crosses between chloramphenicol and erythromycin resistance markers located in mitochondrial DNA demonstrated homoplasmons of parental and recombinant genotypes, and heteroplasmons with recombinant and/or parental genotypes. Although the heteroplasmons were unstable and the homoplasmic components could be segregated by plating on selective media, the heteroplasmic state was often maintained beyond 19 cell divisions when grown on non-selective medium. Homoplasmons of recombinant genotype from repulsion crosses were observed with a frequency of 7.2, 9.0, 11.2 and 11.4 percent; two crosses with the resistance markers in coupling had 5.4 and 11.5 percent recombinants. Under non-selective conditions, the mitochondrial marker derived from the haploid parent of a mating type predominated in zygote-daughter-cells; this asymmetrical distribution could be reversed by selective pressure for the marker transmitted with low frequency. The challenge with chloramphenicol and erythromycin of zygotes from crosses of resistance-markers in repulsion revealed that inter-mitochondrial complementation was not occurring.

Six ethidium bromide (EB) resistant mutants were isolated and characterized. The resistance in three isolates was stable, nuclearly inherited and sensitive to glucose repression. Unstable EB dependent resistance was present in the other isolates. The mutants with stable and one of those with unstable EB resistance showed cross resistance to the cationic detergent, cetyltrimethylammonium bromide. Both anaerobiosis and oligomycin inhibited EB induction of petites in the EB sensitive strain and in the EB-sensitized resistant strains. The observations are discussed in relationship to a mechanism of EB resistance and petite induction at the level of the mitochondrial membrane.

An unusual property of GR25a [rho+] was the production of 20 to 30 percent [rho−] zygote colonies when crossed to a tester strain lacking mitochondrial DNA. Spontaneous [rho−] isolates of GR25a [rho+] were observed to be highly suppressive and to contain mitochondrial DNA of a parental buoyant density (1.685 g/cm3). Three ethidium bromide induced neutral petites of GR25a [rho+] did not have detectable mitochondrial DNA and were neutral in crosses to [rho+] strains. Seven [rho−] zygote colony isolates obtained from crossing GR25a [rho+] to a neutral petite were shown to contain abnormal mitochondrial DNA. Six zygote colony isolates had mitochondrial DNA of a buoyant density less than, or equal to, GR25a (1.682–1.685 g/cm3), whereas one isolate had a buoyant density greater than GR25a (1.688 g/cm3). It was suggested that abnormal mitochondrial DNA is generated during the mating reaction.

The deleterious effects of many environmental agents--ultraviolet (UV) light, ionizing radiation, and polycyclic aromatic hydrocarbons, to cite a few--are primarily attributed to their ability to react with, and thereby structurally modify the deoxyribonucleic acid (DNA) of the living cell1. Given the appreciable level of damage believed to occur naturally in DNA2 and the need to maintain its fidelity as the repository of the genetic script, it s understandable that all living organisms, humans included, should possess multiple cellular processes whose combined actions promote the repair of damage to DNA3. An increasing number of genetically transmitted disorders in man is being found to be associated with enhanced sensitivity to extrinsic DNA-damaging agents. Patients afflicted with any one of these particular Mendelian single-gene traits are typically cancer-prone and suffer from progressive neurodegeneration, and, at least for a few disorders, their cells when cultured in vitro exhibit anomalies in one or more DNA epair processes4–8. The majority of these hereditary diseases can be divided into two broad groups: (i) disorders associated with hypersensitivity to UV light and UV-mimetic chemicals; and (ii) disorders associated with hypersensitivity to ionizing radiation and radiomimetic chemicals. Exemplary diseases of the former group--namely, xeroderma pigmentosum (XP), Bloom syndrome (BS), and Cockayne syndrome--are reviewed elsewhere in this volume. Here we present an overview of the clinical and laboratory features of the latter group of disorders with emphasis on the prototype disease, ataxia telangiectasia (AT)7,8, and comment briefly on current insight into the pathogenesis of these disorders and its implications for theories on environmental carcinogenesis and neurodegenerative processes in man.

We have demonstrated that restriction fragment length polymorphisms (RFLPs) produced by BamHI can be used as markers for constant (C) region heavy chain genes C psi gamma (C gamma pseudogene), C gamma 2, and C gamma 4. These RFLPs were found nonrandomly associated in the population sample studied. Of the eight combinations (haplotypes) of RFLPs theoretically possible, only two accounted for a total of 88% of the 116 chromosomes examined, a value greater than the total of 25% expected from random segregation of alleles. This indicates considerable linkage disequilibrium between C psi gamma, C gamma 2, and C gamma 4. Quantitative assessment of the degree of association between C gamma gene RFLPs, Gm markers, and switch region RFLPs adjacent to C mu and C alpha 1 revealed that C psi gamma is most tightly associated with C gamma 2 (r = 0.81 and 0.95 for the two common haplotypes), suggesting that C psi gamma maps to a position lying between C alpha 1 and C gamma 2. The association analysis used here should have general applicability for studying the genomic organization of other multigene families.

Previous tetrad analyses defined a yeast strain (332-7c) as containing a single nuclear gene (11.8 map units from the centromere) conferring resistance to oligomycin. Resistance to 18 additional inhibitors of mitochondrial function (Table 1) was determined on (i) ascospore isolates from tetrads segregating 2 resistant: 2 sensitive for oligomycin (Table 2) and (ii), spontaneously derived sensitive isolates of the oligomycin resistant strain (Tables 3 and 4). The observed pattern of resistance suggests that the gene for resistance to oligomycin also results in (i) cross resistance to rutamycin, venturicidin, triethyltin bromide, antimycin A, carbonylcyanide m-chlorophenylhydrazone, tetra-N-butylammonium bromide, dibenzyl-dimethylammonium chlorop and tetracycline and (ii), collateral sensitivity to paromomycin, neomycin, dequalinium chloride, ethidium bromide and acriflavin.

We have detected restriction fragment length polymorphisms associated with the immunoglobulin heavy chain C gamma genes. DNA from both parents of an individual having an unbalanced rearrangement of the long arm of chromosome 14, region q32 [Cox, D. W., Markovic, V. D. & Teshima, I. E. (1982) Nature (London) 297, 428-430], revealed distinctive patterns of BamHI fragments which hybridized with cloned probes from the C gamma 2-C gamma 4 gene cluster. The number of hybridizing fragments in both cases (five) equaled the number of known C gamma genes. Pedigree and densitometric analyses indicated that the proband did not have any maternal complement of C gamma gene-hybridizing fragments. Included on the deleted chromosomal segment was a C gamma gene having properties of the previously reported C gamma pseudogene. We also examined DNA from this family with a probe for the highly polymorphic locus D14S1, which recently was demonstrated to be tightly linked to the C gamma 1 gene locus [Balazs, I., Purrello, M., Rubinstein, P., Alhadeff, B. & Siniscalco, M. (1982) Proc. Natl. Acad. Sci. USA 79, 7395-7399]. EcoRI and EcoRI-BamHI fragments from both parents hybridized with a probe for this locus in DNA from the proband, indicating that, unlike the C gamma gene family, D14S1 was not deleted from the abnormal chromosome. Thus, the chromosomal breakpoint in the proband lies within region 14q32 between the two tightly linked markers, D14S1 and the C gamma 1 heavy chain gene locus. The D14S1 locus must lie proximal to the centromere relative to the C gamma gene family. The genetic variability detected with C gamma gene probes may prove useful for genetic analysis of structural rearrangements involving this region of chromosome 14.

Neutral petites were induced in three different [rho+] strains by treating with ethidium bromide. However, EB treatment of spontaneously generated suppressive petites, which were derived from the same [rho+] strains, produced no neutral petites and moreover had little or no effect on the suppressiveness of the petities. Genetic analysis showed that the ethidium bromide resistance of the suppressive petites was a manifestation of cytoplasmically inherited, respiratory deficiency.

When crossed to strain GR25 [rho+], petites lacking in mtDNA were neutral, but when crossed to a related [rho+] strain (GR25a) they were found to be suppressive (Table 1). Likewise, crosses of GR25 [rho+] and GR25a [rho+] to a common [rho+] parent were, respectively, neutral and suppressive (Table 1). The suppressive phenotype observed in these crosses was attributed to a factor in the [rho+] strain GR25a. Strain GR25a also differed from strain GR25 in having a decreased [rho+] stability (Table 2) and a decreased transmission of its cytoplasmically-inherited erythromycin-resistance marker to zygote progeny (Table 4). These three phenotypes of GR25a are, discussed in terms of a nuclear mutation in a gene responsible for the maintenance of the [rho+] state.