Spinal muscular atrophy (SMA) is an autosomal recessive disorder caused by a mutation in SMN1 that stops production of SMN (survival of motor neuron) protein. Insufficient levels of SMN results in the loss of motor neurons, which causes muscle weakness, respiratory distress, and paralysis. A nearly identical gene (SMN2) contains a C-to-T transition which excludes exon 7 from 90% of the mature mRNA transcripts, leading to unstable proteins which are targeted for degradation. Although SMN2 cannot fully compensate for a loss of SMN1 due to only 10% functional mRNA produced, the discovery of the intronic splicing silencer (ISS-N1) opened a doorway for therapy. By blocking its function with antisense oligonucleotides manipulated for high specificity and efficiency, exon 7 can be included to produce full-length mRNA, which then compensates for the loss of SMN1. Nusinersen (Spinraza), the first FDA-approved antisense oligonucleotide drug targeting SMA, was designed based on this concept and clinical studies have demonstrated a dramatic improvement in patients. Novel chemistries including phosphorodiamidate morpholino oligomers (PMOs) and locked nucleic acids (LNAs), as well as peptide conjugates such as Pip that facilitate accurate targeting to the central nervous system, are explored to increase the efficiency of exon 7 inclusion in the appropriate tissues to ameliorate the SMA phenotype. Due to the rapid advancement of treatments for SMA following the discovery of ISS-N1, the future of SMA treatment is highly promising.

The endocannabinoid system is widely studied due to its interactions with cannabis and its role in modulating physiological responses. While most research has focused on the effects of cannabis on adult endocannabinoid systems, recent studies have begun to investigate the role of the endocannabinoid system in developing organisms. However, little is known about the spatial or temporal expression of these receptors during early development. This study combines reverse-transcriptase PCR with in situ hybridizations to compile a timeline of the developmental expression of six key cannabinoid receptors; cb1, cb2, trpv1, trpa1a, trpa1b, and gpr55 in zebrafish embryos, starting from as early as 6 hours post fertilization (hpf) until 3 days post fertilization. This time frame is roughly equivalent to two to ten weeks in human embryonic development. All six genes were confirmed to be expressed within this time range and share similarities with human and rodent expression. Cb1 expression was first detected between 12 and 24 hpf in the retina and CNS, and its expression increased thereafter and was more evident in the olfactory bulb, tegmentum, hypothalamus and gut. Cb2 expression was relatively high at the 6 and 24 hpf timepoints, as determined by RT-PCR, but was undetectable at other times. Trpv1 was first detected at 1 dpf in the trigeminal ganglia, Rohon-Beard neurons and lateral line, and its expression increased in the first 3 days post fertilization (dpf). Expression of trpa1a was first detected as late as 3 dpf in vagal neurons, whereas trpa1b was first detected at 1 dpf associated with trigeminal, glossopharyngeal and vagal ganglia. Expression of gpr55 was diffuse and widespread throughout the brain and head region but was undetectable elsewhere in the embryo. Thus, receptor expression was found to be enriched in the central nervous system and within sensory neurons. This work aims to serve as a foundation for further investigation on the role of cannabinoid and cannabinoid-interacting receptors in early embryonic development.

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