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  • Expression levels of a LAR-like receptor protein tyrosine phosphatase correlate with neuronal branching and arbor density in the medicinal leech.

Expression levels of a LAR-like receptor protein tyrosine phosphatase correlate with neuronal branching and arbor density in the medicinal leech.

Developmental biology (2010-06-15)
Michael W Baker, Eduardo R Macagno
RESUMEN

LAR-like receptor protein tyrosine phosphatases (RPTPs), which are reported to be highly expressed in the nervous systems of most bilaterian animals, have been implicated in the regulation of critical developmental processes, such as neuronal pathfinding, cell adhesion and synaptogenesis. Here we report that two LAR-like RPTPs in the medicinal leech, HmLAR1 and HmLAR2, play roles in regulating the size and density of neuronal arbors within the developing nervous system and in the body wall. Employing single-cell RNAi knockdown and transgene expression techniques, we demonstrate that the expression level of HmLAR1 is directly correlated with the density of an identified neuron's arborization. Knocking down HmLAR1 mRNA levels in the mechanosensory pressure (P) neurons produces a reduced central arbor and a smaller arbor in the peripheral body wall, with fewer terminal branches. By contrast, overexpression of this receptor in a P cell leads to extensive neuronal sprouting, including many supernumerary neurites and terminal branches as well as, in some instances, the normal monopolar morphology of the P cell becoming multipolar. We also report that induced neuronal sprouting requires the expression of the receptor's membrane tethered ectodomain, including the NH(2)-Ig domains, but not of the intracellular phosphatase domains of the receptor. Interestingly, sprouting could be elicited upon ectopic expression of HmLAR1 and the related RPTP, HmLAR2 in the P and other neurons, including those that do not normally express either RPTP, suggesting that the substrates involved in HmLAR-induced sprouting are present in most neurons irrespective of whether they normally express these LAR-like RPTPs. Our data are consistent with the hypothesis that the receptors' ectodomains promote an adhesive interaction that enhances the maintenance of new processes.

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Millipore
Monoclonal Anti-HA−Agarose antibody produced in mouse, clone HA-7, purified immunoglobulin, PBS suspension