Mutations in KATNB1 cause complex cerebral malformations by disrupting asymmetrically dividing neural progenitors.

Exome sequencing analysis of over 2,000 children with complex malformations of cortical development identified five independent (four homozygous and one compound heterozygous) deleterious mutations in KATNB1, encoding the regulatory subunit of the microtubule-severing enzyme Katanin. Mitotic spindle formation is defective in patient-derived fibroblasts, a consequence of disrupted interactions of mutant KATNB1 with KATNA1, the catalytic subunit of Katanin, and other microtubule-associated proteins. Loss of KATNB1 orthologs in zebrafish (katnb1) and flies (kat80) results in microcephaly, recapitulating the human phenotype. In the developing Drosophila optic lobe, kat80 loss specifically affects the asymmetrically dividing neuroblasts, which display supernumerary centrosomes and spindle abnormalities during mitosis, leading to cell cycle progression delays and reduced cell numbers. Furthermore, kat80 depletion results in dendritic arborization defects in sensory and motor neurons, affecting neural architecture. Taken together, we provide insight into the mechanisms by which KATNB1 mutations cause human cerebral cortical malformations, demonstrating its fundamental role during brain development.

A role for juvenile hormone in the prepupal development of Drosophila melanogaster.

To elucidate the role of juvenile hormone (JH) in metamorphosis of Drosophila melanogaster, the corpora allata cells, which produce JH, were killed using the cell death gene grim. These allatectomized (CAX) larvae were smaller at pupariation and died at head eversion. They showed premature ecdysone receptor B1 (EcR-B1) in the photoreceptors and in the optic lobe, downregulation of proliferation in the optic lobe, and separation of R7 from R8 in the medulla during the prepupal period. All of these effects of allatectomy were reversed by feeding third instar larvae on a diet containing the JH mimic (JHM) pyriproxifen or by application of JH III or JHM at the onset of wandering. Eye and optic lobe development in the Methoprene-tolerant (Met)-null mutant mimicked that of CAX prepupae, but the mutant formed viable adults, which had marked abnormalities in the organization of their optic lobe neuropils. Feeding Met(27) larvae on the JHM diet did not rescue the premature EcR-B1 expression or the downregulation of proliferation but did partially rescue the premature separation of R7, suggesting that other pathways besides Met might be involved in mediating the response to JH. Selective expression of Met RNAi in the photoreceptors caused their premature expression of EcR-B1 and the separation of R7 and R8, but driving Met RNAi in lamina neurons led only to the precocious appearance of EcR-B1 in the lamina. Thus, the lack of JH and its receptor Met causes a heterochronic shift in the development of the visual system that is likely to result from some cells 'misinterpreting' the ecdysteroid peaks that drive metamorphosis.

Optomotor-blind of Drosophila melanogaster: a neurogenetic approach to optic lobe development and optomotor behaviour.

The gene optomotor-blind (omb) plays a crucial role in Drosophila optic lobe development. Various mutations in omb lead to different structural defects in the adult optic lobes with correlated behavioural phenotypes. Molecular analysis of omb allows one to trace back behavioural defects to the spatio-temporal misexpression of the gene in mutant development.

Transcript identification in the optomotor-blind locus of Drosophila melanogaster by intragenic recombination mapping and PCR-aided sequence analysis of lethal point mutations.

The optomotor-blind gene of Drosophila melanogaster is large and genetically complex. Five partly independent complementation groups are uncovered by several viable and lethal mutations at the locus. At least 15 RNA signals have been detected by Northern blot analysis. One of them, T3, derived from a 75 kb primary transcript, has been proposed as the carrier of optomotor-blind function, based on the large size of its precursor and its tissue distribution. We here provide direct evidence that T3 is the optomotor-blind transcript. A facile and generally applicable selection scheme for the isolation of intragenic meiotic recombinants was applied to map two lethal optomotor-blind point mutations to exons of the T3 transcript. Amplification of mutant DNA by the polymerase chain reaction (PCR) and sequencing of the amplified exons revealed the presence of mutations that lead to truncation of the T3 open reading frame. The recombination rate observed in the optomoter-blind locus is within the range of rates that have been determined in a few other Drosophila loci.

Mutations in the proximal region of the optomotor-blind locus of Drosophila melanogaster reveal a gradient of neuroanatomical and behavioral phenotypes.

Mutations in the complex optomotor-blind (omb) gene locus (4C4-6) lead to a number of different phenotypes in various tissues of the adult Drosophila melanogaster fly. At the core of the locus lies a lethal complementation group, named l(1)omb, whose mutations cause larval and pupal lethality. Some 40% of all males hemizygous for lethal omb alleles develop to the pharate adult stage. These flies can be rescued from the pupal case and show a severe disturbance in optic lobe development. The recessive viable allele In(1)ombH31 reduces the optomotor response in walking flies and during stationary flight of tethered flies. At the neuroanatomical level, these animals lack a subset of lobula plate giant neurons (LPGNs), which are thought to mediate optomotor behavior. Chromosomal aberrations deleting the proximal, non transcribed part of the locus complement the lethality, but still cause neuroanatomical and optomotor defects. Analysis of different allelic combinations of such mutations, in which increasing amounts of DNA downstream of the transcribed region are removed, reveals a step gradient of increasing severity of the neuroanatomical defects and behavioral phenotypes. On this basis the 3'-regulatory region is divided into three domains each having specific effects on optic lobe development.

Malformations of the regenerating optic tectum of larvae of the Egyptian toad, Bufo regularis Reuss.

Morphological and histological abnormalities were observed in the regenerating optic tecta of Bufo regularis larvae after partial excision of the left tectum and total excision of the right tectum. They were found in both the left and the right tectum. Invagination of the tectal tissue into the optic ventricle, masses of blood capillaries and gaps or cavities in the tectal tissue were observed. The size of the optic tecta was reduced and the shape and structure of the dorsal aspect of the midbrain were highly anomalous.

Mutant alleles at the locus elav in Drosophila melanogaster lead to nervous system defects. A developmental-genetic analysis.

We report a developmental and genetic analysis of the X-linked vital locus l(1)EC7 in Drosophila melanogaster. The locus maps in the salivary band region 1B4-5 to 1B8-9, a part of the X chromosome previously shown to be essential for normal neural development. Certain mutant alleles at the locus can cause embryonic lethality, indicating that the function provided by the gene is essential during embryogenesis. A developmental analysis of gynandromorphic genetic mosaics shows that: (1) the gene function is autonomously essential in the eye; (2) the gene function is essential for normal development of the optic lobes; and (3) the gene function is not necessary in most major imaginal-disc cell derivatives with the exception of the eye disc. Conclusions from the developmental analysis of a temperature sensitive allele are consistent with those from the mosaic analysis. The embryonic lethality caused by the mutant alleles and abnormalities observed in the genetic mosaics have led us to rename the locus l(1)EC7 to elav (embryonic lethal, abnormal visual system).

Neural cell types surviving congenital sensory deprivation in the optic lobes of Drosophila melanogaster.

Golgi staining of neuronal cell types in the optic lobe rudiments of adult eyeless flies of the sine oculis (so) mutant of Drosophila melanogaster reveals partial independence of optic lobe's development from compound eye formation. (1) Differentiation and maintenance of many neuronal cell types of medulla and lobular complex do not require innervation of the medulla from the retina and the lamina. Neurons derived from the outer and inner optic anlage have been found in adult eyeless flies. (2) The rudiments of ipsilateral medulla, lobula, and lobular plate are isotopically connected with each other. (3) Stratification of the lobular complex is retained. (4) Equivalent parts of the dorsal lobulae are connected by heterolateral small field neurons. (5) The shapes of many tangential neurons of the medulla show sprouting and compensatory innervation of the lobular complex. The basic results reported here for eyeless flies have many parallels in what is known about anophthalmic mice.