P40 polyester sheet plastination technique for brain and body slices: The vertical and horizontal flat chamber methods.

The P40 technique produces high-quality brain and body slices and is the user-friendliest of the polyester techniques. The P40 polyester technique follows the same classical steps for plastination. That is, preparation of the specimen, fixation (optional), dehydration by freeze substitution, forced impregnation and curing. Two methods used to prepare two different types of specimens, that is, brain slices and body slices are described. Each method has its own characteristics depending on the specimen type used. Brain slices were used to illustrate the vertical small chamber method while the body slices were used to illustrate the horizontal large chamber method. The brain slices obtained using P40 are of very good quality presenting good contrast between grey and white matter. The body slices are also of very good quality. The physical appearance of these slices makes them an exceptional instrument for diagnostic imaging and anatomical correlation. Body slices prepared with P40 retain the natural colour of the tissue and preserve the anatomical relationships.

Elevated T-maze evaluation of anxiety and memory effects of NMDA/glycine-B site ligands injected into the dorsal periaqueductal gray matter and the superior colliculus of rats.

Rat behaviors in the elevated T-maze (ETM) were evaluated following tectum microinjections of either glycine (GLY, 1, 10, 80 and 120 nmol) or d-serine (D-SER, 160 and 320 nmol), the putative endogenous agonists of GLY-B site at NMDA receptor, or the respective antagonist 7-chloro-kynurenic acid (7CK, 8 nmol). ETM performance was appraised by two validated scores of anxiety, i.e., the inhibitory avoidance duration (AD) and risk assessment behavior, and two scores derived from a newly developed approach to inhibitory avoidance learning curves, i.e., the learning median number of trials (T50) and avoidance variability (standard deviation of learning curve). Effects on aversive memory consolidation were assessed through changes in the AD measured 48 h after the full-acquisition of inhibitory avoidance. Drug effects were compared to those of vehicle. In most cases, microinjection of GLY-B site agonists into the dorsal periaqueductal gray (dPAG) produced increases in AD, which were compatible with an increase in anxiety. However, neither the intra-periaqueductal injection of 80 nmol GLY, nor that of 160 nmol D-SER, increased the AD. On the other hand, these microinjections invariably produced a parallel left shift in avoidance learning curves, thereby reducing the T50 but not the variability. Effects of 120 nmol GLY on AD and T50 were both antagonized by a previous microinjection of 7CK into the dPAG. The inverse relationship of AD and T50 suggests that increases in the anxiety level reduce the number of trials required for the acquisition of inhibitory avoidance. The above data also suggest the higher consistency and drug sensitivity of T50 as compared to the AD. In turn, whereas the microinjection of 120 nmol GLY into the superior colliculus (SC) did not affect the T50, it increased the AD. On the other hand, there was an increase in avoidance variability following the microinjection of either 120 nmol GLY into the SC or 8 nmol 7CK into the dPAG. Therefore, the GLY-B receptors within these structures seem to play opposite roles on avoidance variability. In contrast, neither of these treatments changed T50. Finally, whereas the risk assessment was solely decreased by the microinjection of GLY into the SC, the aversive memory was only impaired by the microinjection of 7CK into the dPAG. Overall, these data suggest that NMDA/GLY-B receptors of dPAG mediate both anxiety and aversive memory, while those in the SC are most likely involved with attention and visuomotor components of risk assessment behavior.