Immunoreactivity of Hu proteins facilitates identification of myenteric neurones in guinea‐pig small intestine

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Abstract
Hu proteins, together with neurone‐specific enolase (NSE), protein gene product 9.5 (PGP‐9.5), microtubule‐associated protein‐2 (MAP‐2) and tubulin beta III isoform, were evaluated immunohistochemically as neuronal markers in whole‐mount preparations and cultures obtained from the myenteric plexus of guinea‐pig small intestine. Anti‐Hu immunostaining marked the ganglion cell somas and nuclei without staining of the neuronal processes in the whole‐mounts and cultures. The ganglion cell bodies were not obscured by staining of multiple neuronal fibres and this facilitated accurate counting of the neurones. MAP2 immunostaining also provided clear images of individual neurones in both whole mounts and cultures. Immunoreactivity for NSE, PGP‐9.5 and tubulin beta III isoform provided sharp images of the ganglion cells in culture, but not in whole‐mount preparations. Strong staining of the neuronal processes in the whole‐mount preparations obscured the profiles of the ganglion cell bodies to such an extent that accurate counting of the total neuronal population was compromised. Anti‐Hu immunostaining was judged to be an acceptable method for obtaining reliable estimates of total numbers of myenteric neurones in relation to other specific histochemical properties such as histamine binding.