Natalia Arias

A half century (1961-2011) of applying microsurgery to experimental liver research.

The development of microsurgery has been dependent on experimental animals. Microsurgery could be a very valuable technique to improve experimental models of liver diseases. Microdissection and microsutures are the two main microsurgical techniques that can be considered for classifying the experimental models developed for liver research in the rat. Partial portal vein ligation, extrahepatic cholestasis and hepatectomies are all models based on microdissection. On the other hand, in portacaval shunts, orthotopic liver transplantation and partial heterotopic liver transplantation, the microsuture techniques stand out. By reducing surgical complications, these microsurgical techniques allow for improving the resulting experimental models. If good experimental models for liver research are successfully developed, the results obtained from their study might be particularly useful in patients with liver disease. Therefore experimental liver microsurgery could be an invaluable way to translate laboratory data on liver research into new clinical diagnostic and therapeutic strategies.

Coupling inflammation with evo-devo.

Inflammation integrates diverse mechanisms that are associated not only with pathological conditions, such as cardiovascular diseases, type 2 diabetes, obesity, neurodegenerative diseases and cancer, but also with physiological processes like reproduction i.e. oogenesis and embryogenesis as well as aging. In the current review we firstly propose that the inflammatory response could recapitulate the phylogenia. In this way, highly conserved inflammatory mechanisms that play a main role in the evolutive development of different animal species, both invertebrates as well as vertebrates, are identified. Therefore, we also hypothesize that inflammation could represent a key tool used by nature to modulate organisms according to the environmental conditions in which these develop. Thus, inflammation could be the pathway by which the environmental factors could be related to the evolutionary development. If so, the diverse human chronic inflammatory diseases that nowadays the Western society suffer would represent the way for adapting to the abrupt changes in their lifestyle. Nonetheless, the distribution of the different pathological conditions varies in terms of intensity and magnitude among Western country populations depending on their genetic polymorphism. In this case, it should be considered that this set of diseases, distributed between all the individuals that constitute the Westernized society, would represent a true Social Inflammatory Syndrome whose final result is its remodeling. In this context, the use of inflammation by the Western society could represent the camouflaged expression of efficient mechanisms of evolution and development. In addition, if the different types of the inflammatory response involved in these diverse chronic pathological conditions could trace the biochemical origins of life, perhaps inflammation could represent an archaeological tool of unsuspected usefulness for understanding our own origin.

Brain metabolism and spatial memory are affected by portal hypertension

Portal hypertension is a major complication of cirrhosis that frequently leads to a neuropsychiatric disorder that affects cognition. The present study was undertaken in order to compare the performance of sham-operated rats (SHAM) and portal hypertension rats (PH) in reference memory tasks in the Morris water maze (MWM). Two groups of animals were used: SHAM group (n = 12) was used as a control group and PH group (n = 12) by the triple portal vein ligation method was used as an animal model of early evolutive phase of PH. The portal pressure was measured in the splenic parenchyma. Our work shows that spatial learning in the MWM is not impaired in PH group although this group showed a one-day delay in the task acquisition compared to the SHAM group. We assessed the brain metabolic activity of the animals by means of cytochrome c-oxidase (COx) histochemistry. Significant changes were found in the CA3, dentate gyrus, basolateral, medial, lateral and central amygdala, showing lower COx activity in the PH group as compared to the SHAM group in all cases. We found no changes in metabolic activity in prefrontal cortex and CA1 area between groups. In fact, different neural networks were shown according to the execution level of the subjects. The early PH evolution induced changes in brain metabolic activity without biggest alterations in spatial memory.

Mapping Metabolic Brain Activity in Three Models of Hepatic Encephalopathy

Cirrhosis is a common disease in Western countries. Liver failure, hyperammonemia, and portal hypertension are the main factors that contribute to human cirrhosis that frequently leads to a neuropsychiatric disorder known as hepatic encephalopathy (HE). In this study, we examined the differential contribution of these leading factors to the oxidative metabolism of diverse brain limbic system regions frequently involved in memory process by histochemical labelling of cytochrome oxidase (COx). We have analyzed cortical structures such as the infralimbic and prelimbic cotices, subcortical structures such as hippocampus and ventral striatum, at thalamic level like the anterodorsal, anteroventral, and mediodorsal thalamus, and, finally, the hypothalamus, where the mammillary nuclei (medial and lateral) were measured. The severest alteration is found in the model that mimics intoxication by ammonia, followed by the thioacetamide-treated group and the portal hypertension group. No changes were found at the mammillary bodies for any of the experimental groups.

Brain network function during shifts in learning strategies in portal hypertension animals

Patients with minimal hepatic encephalopathy exhibit early impairments in their ability to shift attentional set. We employed a task-switching protocol to evaluate brain network changes. Strategy switching requires the modification of both the relevant stimulus dimension and the required memory system. Rats were trained in an allocentric (A) and a cue-guided (C) task using a four-arm maze. To examine priming, we changed the order in which the tasks were presented. Five groups of animals were used: a SHAM (sham-operated) A-C group (n=10), a SHAM C-A group (n=8), a PH (portal hypertension) A-C group (n=8), PH C-A group (n=8), and a naïve group (n=10). The triple portal vein ligation method was used to create an animal model of the early evolutive phase of PH. The animals were tested in the four-arm radial water maze in a single 10-trial session each day for six days (three days for the allocentric task and three days for the cue-guided task). The metabolic activities of the brains were studied with cytochrome oxidase histochemistry, and brain network changes were assessed with principal component analysis. The behavioural results revealed significant increases in the numbers of correct choices across training days in all groups studied, and facilitation of the acquisition of the second task was present in the C-A groups. Moreover, different brain network activities were found; in the experimental groups, the performance of A-C switch involved the prefrontal cortex, and the key structures involved in the C-A switch in the other groups were the dentate gyrus of the dorsal hippocampus and the basolateral and central amygdala. These networks have a common nucleus of structures (i.e., the parietal cortex and the dorsal and ventral striatum), whereas other structures were specifically involved in each type of strategy, suggesting that these regions are part of both circuits and may interact with one another during learning.

Differential contribution of the hippocampus in two different demanding tasks at early stages of hepatic encephalopathy.

The hippocampus has been established as a site of plasticity during the acquisition of spatial memory. The memory for spatial locations is impaired in patients who develop hepatic encephalopathy (HE). We wondered how the hippocampus can manage different hippocampal-dependent tasks in a type B model of the early evolutive phases of HE induced by triple portal vein ligation. We used a one-trial object-place recognition task that involves making judgements about whether a stimulus was encountered before in that location and a reversal learning task performed in the Morris water maze that involves reward-guided behavior and decision making. Our behavioral results showed impairments in the acquisition of both tasks by the portal hypertension group compared with the sham-operated group. To label brain areas related to these tasks, we marked the expression of the c-Fos protein and revealed high c-Fos immunoreactivity in cornu ammonis 1 (CA1), cornu ammonis 3 (CA3) and entorhinal (Ent) cortex of the PH group compared with the SHAM group in the object-place recognition task and a decrease in c-Fos-positive cells in the reversal task in the CA1, CA3, dentate gyrus (DG), cingulate (CG), prelimbic (PL), and infralimbic (IL) cortices in the PH group compared with the SHAM group. In conclusion, the study corroborated the pivotal role of the hippocampus in spatial memory deficits found in the early stages of type B HE and noted its differential contribution in each of the tasks.

The importance of the context in the hippocampus and brain related areas throughout the performance of a fear conditioning task

The importance context has been broadly studied in the management of phobias and in the drug addiction literature. The way in which changes to a context influence behavior after the simple acquisition of a passive avoidance task remains unclear. The hippocampus has long been implicated in the contextual and spatial processing required for contextual fear, but its role in encoding the aversive component of a contextual fear memory is still inconclusive. Our work tries to elucidate whether a change in context, represented as differences in the load of the stimuli, is critical for learning about the context‐shock association and whether this manipulation of the context could be linked to any change in metabolic brain activity requirements. For this purpose, we used an avoidance conditioning task. Animals were divided into three different experimental conditions. In one group, acquisition was performed in an enriched stimuli environment and retention was performed in a typically lit chamber (the PA‐ACQ‐CONTX group). In another group, acquisition was performed in the typically lit chamber and retention was undertaken in the highly enriched chamber (the PA‐RET‐CONTX group). Finally, for the control group, PA‐CN‐CONTX, acquisition, and retention were performed in the enriched stimuli environment. Our results showed that the PA‐ACQ‐CONTX group had longer escape latencies and poorer retention than the PA‐RET‐CONTX and PA‐CN‐CONTX groups after 24 h of acquisition under contextual changes. To study metabolic brain activity, histochemical labelling of cytochrome c‐oxidase (CO) was performed. CO results suggested a neural circuit including the hippocampus, amygdala, thalamus, parahippocampal cortices, and mammillary nuclei that is involved in the learning and memory processes that enable context‐dependent behavior. These results highlight how dysfunction in this network may be involved in the contextualization of fear associations that underlie several forms of psychopathology, including post‐traumatic stress disorder, schizophrenia, and substance abuse disorders.

Brain networks underlying navigation in the Cincinnati water maze with external and internal cues

The present study investigated the behavioural performance and the contributions of different brain regions on a spatial task performed by Wistar rats in the Cincinnati water maze (CWM) in two conditions: one where both distal and proximal visual cues were available (CWM-light group, n=7) and another where visual cues were eliminated by testing in complete darkness (CWM-dark group, n=7). There were differences in the behavioural performance. Energetic brain metabolism revealed significant differences in the infralimbic, orbitofrontal cortex and anterodorsal striatum. At the same time different brain networks were found. The CWM-light group showed a relationship between the orbitofrontal cortex and medial septum, whereas the CWM-dark group revealed three different networks involving the prefrontal cortex, ventral striatum, hippocampus and amygdala nuclei. The study shows that brain activation differs in these two conditions.

The effects of hyperammonemia in learning and brain metabolic activity

Ammonia is thought to be central in the development of hepatic encephalopathy. However, the specific relation of ammonia with brain energy depletions and learning has not been studied. Our work attempts to reproduce an increase in rat cerebral ammonia level, study the hyperamonemic animals’ performance of two learning tasks, an allocentric (ALLO) and a cue guided (CG) task, and elucidate the contribution of hyperammonemia to the differential energy requirements of the brain limbic system regions involved in these tasks. To assess these goals, four groups of animals were used: a control (CHA) CG group (n = 10), a CHA ALLO group (n = 9), a hyperammonemia (HA) CG group (n = 7), and HA ALLO group (n = 8). Oxidative metabolism of the target brain regions were assessed by histochemical labelling of cytochrome oxidase (C.O.). The behavioural results revealed that the hyperammonemic rats were not able to reach the behavioural criterion in either of the two tasks, in contrast to the CHA groups. The metabolic brain consumption revealed increased C.O. activity in the anterodorsal thalamus when comparing the HA ALLO group with the CHA ALLO group. Significant differences between animals trained in the CG task were observed in the prelimbic, infralimbic, parietal, entorhinal and perirhinal cortices, the anterolateral and anteromedial striatum, and the basolateral and central amygdala. Our findings may provide fresh insights to reveal how the differential damage to the brain limbic structures involved in these tasks differs according to the degree of task difficulty.

Sexual metabolic differences in the rat limbic brain

BACKGROUND There is actually limited evidence about the influence of estrogens on neuronal energy metabolism or functional cerebral asymmetry. In order to evaluate this relationship, eight male and sixteen female adult Wistar rats, divided into estrus and diestrus phase, were used to measure basal neuronal metabolic activity in some of the structures involved in the Papez circuit, using cytochrome c oxidase (C.O.) histochemistry. METHOD We used C.O. histochemistry because cytochrome oxidase activity can be considered as a reliable endogenous marker of neuronal activity. RESULTS We found higher C.O. activity levels in diestrus as compared to estrus and male groups in the prefrontal cortex and thalamus. Conversely, neuronal oxidative metabolism was significantly higher in estrus than in diestrus and male groups in the dorsal and ventral hippocampus (CA1 and CA3) and in the mammillary bodies. However, no hemispheric functional lateralization was found in estrus, diestrus or male groups by C.O. activity. CONCLUSIONS These results suggest a modulatory effect of estrogens on neuronal oxidative metabolism.