Neurofilament Heavy Chain Expression Reveals a Unique Parasagittal Stripe Topography in the Mouse Cerebellum

Abstract

Despite the general uniformity in cellular composition of the adult cerebellum (Cb), the expression of proteins such as ZebrinII/AldolaseC and the small heat shock protein HSP25 reveal striking patterns of parasagittal Purkinje cell (PC) stripes. Based on differences in the stripe configuration within subsets of lobules, the Cb can be further divided into four anterior-posterior transverse zones: anterior zone (AZ) = lobules I-V, central zone (CZ) = lobules VI-VII, posterior zone (PZ) = lobules VIII and anterior IX, and the nodular zone (NZ) = lobules posterior IX-X. Here we used whole-mount and tissue section immunohistochemistry to show that neurofilament heavy chain (NFH) expression alone divides all lobules of the mouse Cb into a complex series of parasagittal stripes of PCs. We revealed that the striped pattern of NFH in the vermis of the AZ and PZ was complementary to ZebrinII and phospholipase C ß3 (PLCß3), and corresponded to phospholipase C ß4 (PLCß4). In the CZ and NZ the stripe pattern of NFH was complementary to HSP25 and corresponded to PLCß3. The boundaries of the NFH stripes were not always sharply delineated. Instead, a gradual decrease in NFH expression was observed toward the edges of particular stripes, resulting in domains comprised of overlapping expression patterns. Furthermore, the terminal field distributions of mossy and climbing fibers had a complex but consistent topographical alignment with NFH stripes. In summary, NFH expression reveals an exquisite level of Cb stripe complexity that respects the transverse zone divisions and delineates an intricately patterned target field for Cb afferents.

Neuronal Migration Defect of the Developing Cerebellar Vermis in Substrains of C57BL/6 Mice: Cytoarchitecture and Prevalence of Molecular Layer Heterotopia

“…These searches revealed tissue hybridized for 56 different genes. From this dataset we identified MLH in tissue hybridized for an additional 9 genes with previously reported expression in PCs including: Bai3 [32], Cacna2d2 [33], Clmn [34], Cst3 [35], ERp29 [36], Grid2 [37], Hpca [38], Id2 [39], and Nefh [40]. As was seen for Calb1 and Cacna1g (online suppl.…”

Cerebellar Zones: History, Development, and Function

“…By comparison to zebrin II, some markers have nearly identical patterns, some are mainly complementary, some subdivide zebrin II stripes, and some show stripes where zebrin II does not. In this volume, Roy Sillitoe presents data on the striped expression of NFH in the mouse cerebellum that adds to the known complexity of transverse and sagittal stripe patterns: not only is it unique amongst stripe markers in rodents in showing clear stripes in ALL transverse zones, but these stripes switch their relationships to other stripe markers from transverse zone to transverse zone [12]. In addition, significant overlap is observed between different markers at stripe boundaries, suggesting a much greater degree of molecular complexity than might otherwise be appreciated by any one marker alone.…”

An optimized surgical approach for obtaining stable extracellular single-unit recordings from the cerebellum of head-fixed behaving mice

“…Mouse monoclonal anti-neurofilament heavy (NFH, also called anti-SMI-32; 1:1500) was purchased from Covance (Princeton, NJ, USA). Anti-SMI-32 recognizes the non-phosphorylated form of NFH (see manufacturer product datasheet for details), which on tissue sections labels the soma, dendrites, and axons of adult Purkinje cells, and also basket cell axons and pinceau (Demilly et al, 2011). Rabbit anti-neurogranin (1:500) was raised against full-length recombinant rat neurogranin protein (Chemicon, Temecula, CA, USA; Cat.…”