Krystyna Bogus-Nowakowska

The cocaine- and amphetamine-regulated transcript (CART) immunoreactivity in the amygdala of the pig.

With 5 figures and 1 table

Distribution and chemical coding pattern of the cocaine- and amphetamine-regulated transcript (CART) immunoreactivity in the preoptic area of the pig

This study provides a detailed description of cocaine-and amphetamine-regulated transcript (CART) distribution and the co-localization pattern of CART and gonadotropin releasing hormone (GnRH), somatostatin (SOM), neuropeptide Y (NPY), cholecystokinin (CCK), and substance P (SP) in the preoptic area (POA) of the domestic pig. The POA displays a low density of immunoreactive cells and rich immunoreactivity for CART in fibers. CART-immunoreactive (CART-IR) cell bodies were single and faintly stained, and located in the medial preoptic area (MPA) and the periventricular region of the POA. A high density of immunoreactive fibers was observed in the periventricular preoptic nucleus (PPN); a high to moderate density of fibers was observed in the MPA; but in the dorso-medial region of the MPA the highest density of fibers in the whole POA was observed. The lateral preoptic area (LPA) exhibited a less dense concentration of CART-immunoreactive fibers than the MPA. The median preoptic nucleus (MPN) showed moderate to low expression of staining fibers. In the present study, dual-labeling immunohistochemistry was used to show that CART-IR cell bodies do not contain any GnRH and SP. CART-positive fibers were identified in close apposition with GnRH neurons. This suggests that CART may influence GnRH secretion. Double staining revealed that CART-IR structures do not co-express any of the substances we studied, but a very small population of CART-IR fibers also contain SOM, CCK or SP.

Somatostatin-like immunoreactivity in the amygdala of the pig.

The distribution and morphology of neurons containing somatostatin (SOM) was investigated in the amygdala (CA) of the pig. The SOM-immunoreactive (SOM-IR) cell bodies and fibres were present in all subdivisions of the porcine CA, however, their number and density varied depending on the nucleus studied. The highest density of SOM-positive somata was observed in the layer III of the cortical nuclei, in the anterior (magnocellular) part of the basomedial nucleus and in the caudal (large-celled) part of the lateral nucleus. Moderate to high numbers of SOM-IR cells were also observed in the medial and basolateral nuclei. Many labeled neurons were also consistently observed in the lateral part of the central nucleus. In the remaining CA regions, the density of SOM-positive cell bodies varied from moderate to low. In any CA region studied SOM-IR neurons formed heterogeneous population consisting of small, rounded or slightly elongated cell bodies, with a few poorly branched smooth dendrites. In general, morphological features of these cells clearly resembled the non-pyramidal Golgi type II interneurons. The routine double-labeling studies with antisera directed against SOM and neuropeptide Y (NPY) demonstrated that a large number of SOM-IR cell bodies and fibers in all studied CA areas contained simultaneously NPY. In contrast, co-localization of SOM and cholecystokinin (CCK) or SOM and vasoactive intestinal polypeptide (VIP) was never seen in cell bodies and fibres in any of nuclei studied. In conclusion, SOM-IR neurons of the porcine amygdala form large and heterogeneous subpopulation of, most probably, interneurons that often contain additionally NPY. On the other hand, CCK- and/or VIP-IR neurons belonged to another, discrete subpopulations of porcine CA neurons.

The densities of calbindin and parvalbumin, but not calretinin neurons, are sexually dimorphic in the amygdala of the guinea pig

In the amygdala, the calcium-binding proteins (calbindin, parvalbumin or calretinin) are useful markers of specific subpopulations of γ-aminobutyric acid (GABA) containing neurons. In the rat and monkey they together mark the vast majority of GABA-containing neurons in this brain region. As GABA involvement in the control of various behaviors in a sex-specific manner and sexual dimorphism of the GABAergic system itself were recently proven, the question is how much dimorphic may be various subpopulations of this system. Thus, the present study investigates for the first time the presence/absence of sexual dimorphism among neurons expressing calbindin (CB), parvalbumin (PV) and calretinin (CR) which form in the amygdala main subsets of GABAergic system. The results show that in the amygdala of the guinea pig the densities of CB and/or PV expressing neurons are sexually dimorphic with the female>male pattern of sex differences in the basolateral amygdala. In the medial and cortical amygdala respectively CB and PV values are also sexually dimorphic, favoring males. The densities of CR expressing neurons are in the amygdala of the guinea pig sexually isomorphic. In conclusion, the results of the present study provide an evidence that in the amygdala of the guinea pig the densities of neurons expressing CB and/or PV are sexually dimorphic what supports the idea that GABA participates in the mediation of sexually dimorphic functions, controlled by this brain area.

Embryonic and postnatal development of calcium-binding proteins immunoreactivity in the anterior thalamus of the guinea pig.

Our recent studies have shown that the distribution of calretinin (CR) in the anterior thalamic nuclei (ATN) changes significantly during the development of the guinea pig. The present study was designed to reveal the distribution pattern of calcium-binding proteins, i.e. calbindin (CB) and parvalbumin (PV), as well as the colocalization pattern of all three proteins, including CR, in the ATN of guinea pigs ranging from the 40th embryonic day (E40) to the 80th postnatal day (P80). According to these patterns, CB appears exclusively in the perikarya of the anteromedial nucleus (AM) not before P20 and always colocalizes with CR. Moreover, CB and CR colocalize in fibers of thin bundles traversing the anteroventral nucleus (AV) since E50. The ATN also display CB-positive neuropil in all studied stages, especially a strong one in the ventral part of the AV. PV was not observed in the perikarya of the ATN in all the stages, but was abundantly present in the neuropil of the anterodorsal nucleus (AD). No colocalizations exist between PV and the rest of the studied proteins. In conclusion, our study reveals that the distribution of the studied proteins differs greatly. Nevertheless, the postnatal coexistence of CB and CR in the AM perikarya may indicate the cooperation of both of the proteins in some functions of the nucleus. Parvalbumin is limited mostly to the neuropil of the AD, suggesting different functions in comparison to CB and CR.

The cocaine- and amphetamine-regulated transcript, calbindin, calretinin and parvalbumin immunoreactivity in the medial geniculate body of the guinea pig.

The purpose of this study was to describe the distribution and colocalization of cocaine- and amphetamine-regulated transcript (CART) and three calcium-binding proteins (calbindin, calretinin and parvalbumin) in each main division of the medial geniculate body (MGB) in the guinea pig. From low to moderate CART immunoreactivity was observed in all divisions of the MGB, although in most of its length only fibers and neuropil were labeled. A small number of CART immunoreactive somata were observed in the caudal segment of the MGB. The central parts of all divisions contained a distinctly smaller number of CART immunoreactive fibers relative to their outer borders, where CART fibers formed patchy clusters. As a whole, the intense CART immunoreactive borders formed a shell around the weakly CART labeled core. Double-labeling immunofluorescence showed that CART did not colocalize with either calbindin, calretinin or parvalbumin, whose immunoreactivity was predominantly restricted to perikarya. The distribution pattern of calretinin was more similar to that of calbindin than to that of parvalbumin. Calretinin and calbindin exhibited higher immunoreactivity in the medial and dorsal divisions of the MGB, where parvalbumin staining was low. In general, although parvalbumin exhibited the weakest immunoreactivity of all studied Ca(2+) binding proteins, it was most highly expressed in the ventral division of the MGB. Our results indicate that CART could be involved in hearing, although its immunoreactivity in the medial geniculate complex was not as intense as in other sensory brain regions. In the guinea pig the heterogeneous and complementary pattern of calbindin, calretinin and parvalbumin is evident, however, the overlap in staining appears to be more extensive than that seen in other rodents.

Cocaine- and amphetamine-regulated transcript and calcium binding proteins immunoreactivity in the subicular complex of the guinea pig.

In this study we present the distribution and colocalization pattern of cocaine- and amphetamine-regulated transcript (CART) and three calcium-binding proteins: calbindin (CB), calretinin (CR) and parvalbumin (PV) in the subicular complex (SC) of the guinea pig. The subiculum (S) and presubiculum (PrS) showed higher CART-immunoreactivity (-IR) than the parasubiculum (PaS) as far as the perikarya and neuropil were concerned. CART- IR cells were mainly observed in the pyramidal layer and occasionally in the molecular layer of the S. In the PrS and PaS, single CART-IR perikarya were dispersed, however with a tendency to be found only in superficial layers. CART-IR fibers were observed throughout the entire guinea pig subicular neuropil. Double-labeling immunofluorescence showed that CART-IR perikarya, as well as fibers, did not stain positively for any of the three CaBPs. CART-IR fibers were only located near the CB-, CR-, PV-IR perikarya, whereas CART-IR fibers occasionally intersected fibers containing one of the three CaBPs. The distribution pattern of CART was more similar to that of CB and CR than to that of PV. In the PrS, the CART, CB and CR immunoreactivity showed a laminar distribution pattern. In the case of the PV, this distribution pattern in the PrS was much less prominent than that of CART, CB and CR. We conclude that a heterogeneous distribution of the CART and CaBPs in the guinea pig SC is in keeping with findings from other mammals, however species specific differences have been observed.

Distribution of the Cocaine and Amphetamine - Regulated Transcript (Cart) and Calcium Binding Proteins Immunoreactivity in the Preoptic Area of the Ram

Abstract The aim of the study was to describe the distribution of cocaine and amphetamine-regulated transcript (CART) and calcium binding proteins (CaBPs) of EF-hand family, namely calbindin, calretinin, and parvalbumin in the preoptic area (POA) of the ram. Frozen sections were processed for a routine immunofluorescence labelling. CART, calbindin, and calretinin immunoreactivity was present in neurons and fibers of the preoptic area, whereas parvalbumin showed immunoreactivity only in the POA fibers. CART displayed from a moderate to low immunoreactivity in cells and a high immunoreactivity in fibers. The highest immunoreactivity of all studied CaBPs exhibited calbindin, whereas the lowest parvalbumin. The results of the present study suggest that among the studied CaBPs, calbindin is the most likely to be involved in the participation of the important regulatory functions in the ram’s POA and the rich CART innervation seems to be strictly related to its control of the reproduction.

A Morphometric Comparative Study of the Medial Geniculate Body of the Rabbit and the Fox

With 6 figures and 2 tables

Distribution of Galanin and Galanin Receptor 2 in the Pre‐optic Area of the Female Guinea Pig

This study describes the distribution of galanin (Gal) and galanin receptor 2 (GalR2) in the pre‐optic area (POA) of the female guinea pig. Frozen sections were undergone for a routine immunofluorescence labelling. Gal and GalR2 display immunoreactivity in all parts of the pre‐optic area. Gal shows reactivity both in perikarya and fibres, whereas GalR2 was observed only in perikarya. Gal‐ and GalR2‐immunoreactive (‐ir) perikarya were the most numerous in the medial pre‐optic area (MPA) with the highest reactivity in its dorsal part. In the median pre‐optic nucleus (MPN) and periventricular pre‐optic nucleus (PPN), only single Gal‐ and GalR2‐ir neurons were observed. The highest density of Gal‐ir fibres was revealed in the PPN and the lowest in the lateral pre‐optic area (LPA). The results of this study indicate that the distribution pattern of Gal containing neurons overlaps well with the distribution pattern of GalR2‐positive neurons, especially in the MPA. This may suggest GalR2‐dependent activity in this brain region.