Differential regulation of substance P and somatostatin in Martinotti cells of the developing cat visual cortex

Abstract

In order to determine their morphological development and ontogenetic fate, Martinotti neurons immunoreactive for substance P and somatostatin have been analysed in the cat visual cortex. Martinotti neurons are located in layers V and VI. They are multipolar to bitufted, and most dendrites remain in layers V and VI. Their typical features is the ascending axon, which emerges from an apical dendrite or from the upper pole of the soma. A number of collaterals branch off in layer V, forming a local terminal plexus. The axon then branches into 2-8 collaterals, which ascend as a bundle to layers III and II, where a second terminal plexus is formed. Some collaterals ascend to layer I where they adopt a horizontal course. Horizontal collaterals in the terminal layers V, III, II, and in layer I may reach up to 400 microns in length. Martinotti neurons begin to differentiate perinatally. The quantitative analysis reveals that the initial time course of differentiation of Martinotti cells is very similar in material stained for substance P and for somatostatin. Double immunofluorescence then confirms that the two peptides are colocalized in Martinotti cells of layers V and VI during the early postnatal period. Further, substance P is colocalized with GABA. Substance P expression in Martinotti cells can be observed only in the immature visual cortex. After postnatal day 15, the Martinotti neuron system becomes less and less detectable by substance P immunoreactivity. It declines to virtually undetectable levels after the third postnatal month. The adult visual cortex is almost devoid of substance P-immunoreactive cell bodies, processes and axon terminals. In situ hybridization confirms this finding, revealing beta-preprotachykinin mRNA-expressing cell bodies in layers V and IV at postnatal day (P)6 and P12, but not in the adult cortex. This suggests a downregulation of the substance P expression at the transcriptional level. In contrast, somatostatin-immunoreactive Martinotti cells, most of which have coexpressed substance P during early postnatal life, can still be observed in the adult cortex. Thus, the Martinotti neurons constitute a persisting cell type, although many individual neurons of this type disappear during the second postnatal month by degeneration and cell death. In summary, while somatostatin is permanently expressed in Martinotti neurons in the cat visual cortex, substance P peptide and mRNA are transiently expressed during an early postnatal period, and apparently are downregulated later in development.