Selective vulnerability in neuronal populations in nmd/SMARD1 mice.

Spinal muscular atrophy with respiratory distress type 1 (SMARD1) is an autosomal recessive motor neuron disease causing distal limb muscle atrophy that progresses proximally and is accompanied by diaphragmatic paralysis. Neuromuscular junction (NMJ) alterations have been reported in muscles of SMARD1 model mice, known as nmd mice, with varying degrees of severity, suggesting that different muscles are specifically and selectively resistant or susceptible to denervation. To evaluate the extent of NMJ pathology in a broad range of muscles, a panel of axial and appendicular muscles were isolated and immunostained from nmd mice. These analyses revealed that selective distal appendage muscles were highly vulnerable to denervation. Susceptibility to pathology was not limited to NMJ alterations, but included defects in myelination within those neurons innervating susceptible muscles. Interestingly, end plate fragmentation was present within all muscles independent of the extent of NMJ alterations, suggesting that end plate fragmentation is an early hallmark of SMARD1 pathogenesis. Expressing the full-length IGHMBP2 cDNA using an adeno-associated virus (AAV9) significantly decreased all aspects of muscle and nerve disease pathology. These results shed new light onto the pathogenesis of SMARD1 by identifying specific motor units that are resistant and susceptible to neurodegeneration in an important model of SMARD1.

Optimization of Morpholino Antisense Oligonucleotides Targeting the Intronic Repressor Element1 in Spinal Muscular Atrophy.

Loss of Survival Motor Neuron-1 (SMN1) causes Spinal Muscular Atrophy, a devastating neurodegenerative disease. SMN2 is a nearly identical copy gene; however SMN2 cannot prevent disease development in the absence of SMN1 since the majority of SMN2-derived transcripts are alternatively spliced, encoding a truncated, unstable protein lacking exon 7. Nevertheless, SMN2 retains the ability to produce low levels of functional protein. Previously we have described a splice-switching Morpholino antisense oligonucleotide (ASO) sequence that targets a potent intronic repressor, Element1 (E1), located upstream of SMN2 exon 7. In this study, we have assessed a novel panel of Morpholino ASOs with the goal of optimizing E1 ASO activity. Screening for efficacy in the SMNΔ7 mouse model, a single ASO variant was more active in vivo compared with the original E1(MO)-ASO. Sequence variant eleven (E1(MOv11)) consistently showed greater efficacy by increasing the lifespan of severe Spinal Muscular Atrophy mice after a single intracerebroventricular injection in the central nervous system, exhibited a strong dose-response across an order of magnitude, and demonstrated excellent target engagement by partially reversing the pathogenic SMN2 splicing event. We conclude that Morpholino modified ASOs are effective in modifying SMN2 splicing and have the potential for future Spinal Muscular Atrophy clinical applications.

Metagenome-Assembled Genome Sequence of Dolichospermum circinale Strain Clear-D4, Assembled from a Harmful Cyanobacterial Bloom Enrichment Culture.

Dolichospermum circinale (formerly Anabaena circinale) is a significant harmful algal bloom species. We report the draft metagenome-assembled genome (MAG) for a strain of D. circinale (Clear-D4) obtained from an enrichment culture. The genome sequence comprises 5,029,933 bp in 560 contigs with a GC content of 37%.

Metagenome-Assembled Genome Sequence of Kapabacteriales Bacterium Strain Clear-D13, Assembled from a Harmful Algal Bloom Enrichment Culture.

Metagenomic sequencing of a Dolichospermum circinale enrichment culture resulted in the assembly of several cocultured metagenome-assembled genomes (MAGs). One MAG was affiliated with the class Kapabacteriales and included 5,724,991 bp in 127 contigs with a GC content of 48.4%.

Metagenome-Assembled Genome Sequence of Rhodobacteraceae Bacterium Strain Clear-D3, Assembled from a Dolichospermum Consortium from Clear Lake, California.

Here, we report the metagenome-assembled genome sequence of a Rhodobacteraceae bacterium strain, Clear-D3, that was reconstructed from a cyanobacterial enrichment from a eutrophic lake. The draft genome sequence shows evidence of an anoxygenic photoautotrophic lifestyle. Other potential capabilities include aerobic heterotrophy, flagellar motility, chemotaxis, and utilization of complex C-P compounds.

Metagenome-Assembled Genome Sequence of Vulcanococcus sp. Strain Clear-D1, Assembled from a Cyanobacterial Enrichment Culture.

We report the metagenome-assembled genome sequence of a Vulcanococcus sp. binned from a cyanobacterial enrichment culture. The genome contains 39 contigs comprising 2.96 Mbp and is estimated as 100% complete, with a GC content of 63.9% and 3,261 predicted coding genes.

Metagenome-Assembled Genome Sequence of Aphanizomenon flos-aquae Strain Clear-A1, Assembled from an Enrichment Culture.

Aphanizomenon flos-aquae is a significant harmful algal bloom-forming cyanobacterial species. Here, we report the draft genome for a strain of A. flos-aquae (Clear-A1) from a harmful algal bloom enrichment culture. This metagenome-assembled genome (MAG) sequence comprises 4,452,466 bp in 60 contigs with a GC content of 37.1%.