Ultrastructural localization of tyrosine hydroxylase immunoreactivity in the rat diagonal band of broca
Authors: Milner T. A. 
Published: January 2005

The present study sought to establish the cellular basis for the catecholaminergic (i.e., noradrenaline and dopamine) modulation of neurons in the horizontal limb of the diagonal band of Broca (HDB) in the rat brain. The light and electron microscopic localization of antigenic sites for a polyclonal antibody directed against the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH), were examined in the HDB using a double‐bridged, peroxidase‐antiperoxidase method. By light microscopy, numerous punctate, varicose processes with intense TH‐immunoreactivity (TH‐I) were detected in the HDB. Additionally, a few small, bipolar, or multipolar TH‐immunoreactive neurons were observed.

Ultrastructural analysis of single sections revealed that the TH‐labeled processes were axons and axon terminals. Axons (n = 134) with TH‐I were primarily unmyelinated. Terminals with TH‐I (n = 169) were 0.3–1.4 μm in diameter and contained many small, clear vesicles and 0–5 larger dense‐core vesicles. The types of associations (i.e., asymmetric synapses, symmetric synapses, and appositions which lacked a membrane specialization in the plane of section analyzed) formed by the TH‐labeled terminals were quantitatively evaluated. The TH‐labeled terminals: (1) formed associations with unlabeled perikarya and dendrites (134 out of 169), (2) were closely apposed without glial intervention to unlabeled and TH‐labeled terminals (11 out of 169), or (3) had no neuronal associations in the plane of section analyzed (24 out of 169). The relatively rare (n = 4) associations with unlabeled perikarya were mostly characterized by symmetric synaptic specializations. The majority of the TH‐labeled terminals were associated with the shafts of small dendrites (66% of 134). Moreover, most of the associations on dendrites and dendritic spines were further characterized by asymmetric synaptic specializations; however, many were also appositions without any apparent glial intervention in the plane of section analyzed. Additionally, the TH‐labeled terminals were often associated with only one dendrite, which, in the same plane of section, was sparsely innervated by other terminals. Astrocytic processes usually surrounded the portions of the terminals and dendrites not involved in the region of association.

The TH‐immunoreactive perikarya were small (7–12 μm), ovoid, and had an indented nucleus with some heterochromatin. Their scant cytoplasm contained mitochondria, Golgi complexes, and endoplasmic reticulum. A few immunoreactive dendrites, presumably derived from the local neurons, were also detected. Both TH‐immunoreactive perikarya and dendrites were associated primarily with unlabeled terminals, although a few terminals with TH‐I also contacted them.

These results provide ultrastructural evidence showing that in the HDB many TH‐immunoreactive processes are terminals that form direct synapses on unlabeled and a few TH‐labeled neurons and more rarely are in apposition with other unlabeled and labeled terminals. Thus they may provide a cellular substrate for both pre‐ and postsynaptic modulation of septal function by norepinephrine and dopamine, the major catecholamines in this region.