Defense Date

2011

Document Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Neuroscience

First Advisor

William Guido

Abstract

The dorsal lateral geniculate nucleus (dLGN) has become an important model for studying many aspects of visual system development. To date, studies have focused on the development of retinal projections and the role of activity in shaping the pattern of synaptic connections made with thalamocortical relay cells. By contrast, little is known about relay cells and the factors that regulate the growth and establishment of their dendritic architecture. In many systems, such growth seems consistent with the synaptotrophic hypothesis which states that synapse formation and dendritic growth work in a concerted fashion such that afferent input and the establishment of functional synapses are needed to shape the maturation of dendritic arbors. To address this, we characterized the development of relay cells in the dLGN of wild-type (WT) mouse. By adopting a loss of function approach, we assessed the manner in which growth and maturation of relay cells were affected by retinal innervation. For this, we made use of the math-null (math5-/-) mouse in which progenitors fail to differentiate into retinal ganglion cells (RGCs), and exhibit a >95% cell loss. Anterograde labeling of RGC axons with cholera toxin subunit B (CTB), immunolabeling of RGC-specific presynaptic machinery in dLGN (e.g. vesicular glutamate transporter 2), and ultrastructural analysis at the electron microscopy level demonstrated that the dLGN is devoid of retinal innervation. We examined the functional and morphological characteristics of relay cells in WT and math5-nulls during early postnatal life by conducting in vitro whole cell recordings in slices containing dLGN. Individual relay cells were labeled by intracellular injection of biocytin, and imaged by confocal microscopy to obtain the 3-D reconstructions of their dendritic trees. Morphometric analysis revealed that relay cells in WT undergo two growth spurts: an early one where cell class specification and dendritic complexity are established and a later one marked by an increase in dendritic field and length. Following the third week, relay cells growth was stabilized. In math5-nulls, relay cells maintained their morphological identity whereby cells could be classified in three groups (Y: spherical, X: bi-conical, W: hemi-spherical). However, the dLGN was highly reduced in size, and relay cells showed disrupted growth spurts. Relay cells had smaller somata and exhibited fluctuations in dendritic complexity and field extent compared to age-matched WTs. Exuberance in dendritic branching was noted in week 2, and by week 5, relay cells had significantly smaller surface area resulting from a loss of dendritic segments and a reduction in dendritic field extent. Control experiments using RT-PCR revealed that these changes were not due to the loss of math5 in the dLGN. Whole cell recordings and voltage responses to square wave current pulses showed that math5-nulls possess the full compliment of intrinsic membrane properties, such as relay cells displayed both burst and tonic firing modes. A cross of the math5-null with a transgenic mouse that expresses GFP in layer VI cortical neurons revealed a dense plexus of corticogeniculate terminals throughout the mature dLGN. However, the rate of corticogeniculate innervation was highly accelerated and was complete a week earlier than WT. Electric stimulation of cortical axons revealed that synapses are functional and responses were indistinguishable from WT. Taken altogether, these observations suggest that retinal innervation plays an important trophic role in the maturation of dLGN and is necessary for the continued maintenance of relay cells’ structural integrity. However, the general form and function of relay cells seem largely unaffected by the loss of retinal innervation.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

September 2011

Included in

Neurosciences Commons

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