The sea lamprey recovers normal-appearing locomotion after spinal cord transection and its spinal axons regenerate selectively in their correct paths. However, among identified reticulospinal neurons some are consistently bad regenerators and only about 50% of severed reticulospinal axons regenerate through the site of injury. We previously suggested (Shifman and Selzer, 2000) that selective chemorepulsion might explain why some neurons are bad regenerators and others not. To explore the role of additional chemorepulsive axonal guidance molecules during regeneration, we examined the expression of the repulsive guidance molecule (RGM) and its receptor neogenin by in situ hybridization and quantitative PCR. RGM mRNA was expressed in the spinal cord, primarily in neurons of the lateral gray matter and in dorsal cells. Following spinal cord transection, RGM message was downregulated in neurons close (within 10 mm) to the transection at 2 and 4 weeks, although it was upregulated in reactive microglia at 2 weeks post-transection. Neogenin mRNA expression was unchanged in the brainstem after spinal cord transection, and among the identified reticulospinal neurons, was detected only in 鈥渂ad regenerators, Neurons that are known to regenerate well never expressed neogenin. The downregulation of RGM expression in neurons near the transection may increase the probability that regenerating axons will regenerate through the site of injury and entered caudal spinal cord.
The virulence of seven geographic isolates of white spot syndrome virus (WSSV; genus Whispovirus; China [strain CH1995], Nicaragua [strain N2000], Honduras [strain H2000], Ecuador [strains E-L1999 and E-LT2002], and Mexico [strains M-M2001 and M-LP2001]) was compared using a series of challenge experiments, each lasting 10 d. For each isolate, four quantified dilutions (10(-6), 10(7), 10(-8), and 10(-9)) of a viral inoculum were prepared from WSSV-infected shrimp tissue. Each viral inoculum was injected into 10 specific pathogen-free juvenile Pacific white shrimp Litopenaeus vannamei (0.25-1.50 g); controls received injections of marine crustacean physiological saline (3.2%). The minimum dose of viral inoculum that killed 50% of injected shrimp (LD50) was calculated for dilution, tissue concentration, and viral DNA amount. The CH1995 and M-M2001 isolates were the least virulent, with LD50 values of 10(-6) to 10(-7) of viral inoculum. The isolates could be grouped into three virulence clusters (CH1995 and M-M2001; N2000 and E-LT2002; and H2000, E-L1999, and M-LP2001). Virulence clusters were not altered by LD50 values based on viral DNA concentration, although a slight shifting of order in regards to virulence was seen among the three most virulent isolates (E-L1999, H2000, and M-LP2001). Overall, results indicate that there is a measurable virulence difference among WSSV isolates, which may correspond to geographical region.
In mammals, there are three neurofilament (NF) subunits (NF-L, NF-M, and NF-H), but it was thought that only a single NF, NF180, exists in lamprey. However, NF180 lacked the ability to self-assemble, suggesting that like mammalian NFs, lamprey NFs are heteropolymers, and that additional NF subunits may exist. The present study provides evidence for the existence of a lamprey NF-L homolog (L-NFL). Genes encoding two new NF-M isoforms (NF132 and NF95) also have been isolated and characterized. With NF180, this makes three NF-M-like isoforms. In situ hybridization showed that all three newly cloned NFs are expressed in spinal cord neurons and in spinal-projecting neurons of the brainstem. Like NF180, there were no KSP multiphosphorylation repeat motifs in the tail regions of NF132 or NF95. NF95 was highly identical to homologous parts of NF180, sharing 2 common pieces of DNA with it. Northern blots suggested that NF95 may be expressed at very low levels in older larvae. The presence of L-NFL in lamprey CNS may support the hypothesis that as in mammals, NFs in lamprey are obligate heteropolymers, in which NF-L is a required subunit.
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