A novel preclinical rodent model of collagenase-induced germinal matrix/intraventricular hemorrhage.

Germinal matrix/intraventricular hemorrhage (GMH/IVH) is a complication that arises in premature infants associated with neurological sequelae. Greater understanding of GMH/IVH is needed to develop therapies, a goal that depends on the existence of appropriate animal models. Towards this goal, we aimed to develop a rodent model of GMH/IVH based on collagenase-induced hemorrhage that exhibits histological and neurological consequences similar to that seen in patients. Male 6-day-old rats were placed on a warming pad and anesthetized with halothane/nitrous oxide delivered by face mask. Uni- or bilateral periventricular injections of 2-µl collagenase (2.0 U) were performed freehand with a needle inserted percutaneously. Sham rats were infused with saline. Early neonatal development, long-term motor and cognitive performances and alterations in brain volume were assessed. Collagenase-based GMH/IVH negatively affected ambulation, surface righting and negative geotaxis outcomes more evidently in bilaterally infused rats, which also presented an early decrease in brain volume, as assessed by the Cavalieri method. In adult animals, a unilateral collagenase infusion produced no significant alteration on forepaw preference. Only bilaterally infused rats presented an impairment of object recognition memory and locomotor deficit. Nevertheless, histological evaluation also demonstrated a persistent brain volume reduction in bilaterally infused rats. Our study provides a pioneering animal model of collagenase-based GMH/IVH, which can be used to evaluate preventive strategies and potential therapeutic interventions for this disorder.

Acetylcholinesterase activity in the human subfornical organ.

The acetylcholinesterase activity (AChE) in the human subfornical organ (SFO) was detected by the method of Koelle and Friedenwald in 16 human brains collected between 6 and 12 hrs postmortem. The only AchE-positive structures found were neuronal cell bodies and processes, morphologically classified as stellate and fusiform neurons of large, medium and small size. Large ones prevailed in the dorsal zone. The neurons were homogeneously distributed in the rostral area of the SFO. The penetration and ramification of large blood vessels produced a decreasing neuronal density in the medium-caudal area. The architecture of the SFO in sagittal sections comprised a central zone with neurons juxtaposed to the walls of the vascular plexus, whose dendritic and axonal processes showed an intricate pattern without a special arrangement. This neuronal zone of the SFO was surrounded by a peripheral layer of neurons with axonal projections to the rostral area. This layer was thicker in the dorsal zone of the SFO, where axonal fibers "climbing-up" from the central perivascular neurons could be demonstrated. In coronal sections some neurons with prolongations of arcuate distribution connected the dorsal and ventral zones.