GABAergic and Glutamatergic Phenotypes of Neurons Expressing Calcium-Binding Proteins in the Preoptic Area of the Guinea Pig.

The mammalian preoptic area (POA) has large populations of calbindin (CB), calretinin (CR) and parvalbumin (PV) neurons, but phenotypes of these cells are unknown. Therefore, the question is whether neurons expressing CB, CR, and/or PV are GABAergic or glutamatergic. Double-immunofluorescence staining followed by epifluorescence and confocal microscopy was used to determine the coexpression patterns of CB, CR and PV expressing neurons with vesicular GABA transporters (VGAT) as specific markers of GABAergic neurons and vesicular glutamate transporters (VGLUT 2) as specific markers of glutamatergic neurons. The guinea pig was adopted as, like humans, it has a reproductive cycle with a true luteal phase and a long gestation period. The results demonstrated that in the guinea pig POA of both sexes, ~80% of CB+ and ~90% of CR+ neurons coexpress VGAT; however, one-fifth of CB+ neurons and one-third of CR+ cells coexpress VGLUT. About two-thirds of PV+ neurons express VGAT, and similar proportion of them coexpress VGLUT. Thus, many CB+, CR+ and PV+ neurons may be exclusively GABAergic (VGAT-expressing cells) or glutamatergic (VGLUT-expressing cells); however, at least a small fraction of CR+ cells and at least one-third of PV+ cells are likely neurons with a dual GABA/glutamate phenotype that may coexpress both transporters.

2,3,7,8-Tetrachlorodibenzo-p-dioxin alters steroid secretion but does not affect cell viability and the incidence of apoptosis in porcine luteinised granulosa cells.

The compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), a by-product of human industrial activity, was found to affect ovarian steroidogenesis in animals, but the mechanism of its action is still unclear. The aims of the study were to examine the effect of TCDD on (1) progesterone (P4) and oestradiol (E2) production by granulosa cells isolated from medium (3-6 mm) and preovulatory (≥ 8 mm) porcine follicles, (2) the viability of the cells, and (3) the incidence of apoptosis. Porcine granulosa cells were cultured (48 h) with or without TCDD (100 pM, 100 nM). Steroid hormone concentrations in the medium were determined by radioimmunoassay. The viability of granulosa cells was tested spectrophotometrically (alamarBlue™ assay). Apoptosis was evaluated by flow cytometry using Annexin V and by TUNEL assay. The higher dose of TCDD (100 nM) significantly inhibited P4 and stimulated E2 production by luteinised granulosa cells isolated from medium follicles. The lower dose of TCDD (100 pM) significantly stimulated P4 and inhibited E2 secretion by the cells isolated from preovulatory follicles. None of the two TCDD doses affected cell viability or induced apoptosis in granulosa cells. In conclusion, TCDD directly affected steroid production by granulosa cells obtained from mature pigs, but the effect of TCDD was not due to its cytotoxicity.

Advancing the impact identification step of benefit-cost analysis of potable water infrastructure investments: A systems method for identifying important impacts pre-monetisation.

Benefit cost analysis (BCA) is frequently used to evaluate potable water infrastructure (PWI) investments. However, a limitation raised by BCA researchers is the narrow view of analysts in identifying investment impacts. In this paper, we propose a systems-thinking framework, supported by data from the literature, interviews, and macroeconomic data, to provide analysts with a more systematic and comprehensive view of investment impacts. The framework, once built, can be applied to any PWI investment question, to identify the prominent impacts that an analyst should consider taking forward through the quantification stages of the BCA process. We validate our method for identifying impacts using data from New Zealand. Our method identifies impacts that are typically not valued in BCA of PWI investments, but that are a common impact of many types of PWI investment decision. Household costs, for example, score in the Top 10 investment outcomes, but are only typically valued in ex post analyses of outbreaks. These impacts warrant attention in future benefit cost analyses. An additional contribution is the development a new betweenness importance rating, which we call flow betweenness, to evaluate each impact's prominence within the PWI socio-economic system.

Update on prevalence of Babesia canis and Rickettsia spp. in adult and juvenile Dermacentor reticulatus ticks in the area of Poland (2016-2018).

Ornate dog tick, Dermacentor reticulatus is an important vector of Babesia canis, and Rickettsia spp. and other pathogens of veterinary and public health interest. The current study is the first to investigate the long-term changes in prevalence of these pathogens in expanding tick populations in Central Europe. Molecular techniques (PCR, sequencing) were applied for the detection of pathogen DNA in adult (n = 2497) and juvenile ticks (1096 larvae and 410 nymphs). DNA of Rickettsia spp. was identified in 35% of adults and 12.6% of juvenile ticks. DNA of B. canis was detected in 3% of adult ticks and only in ticks from the Eastern region (regional prevalence 6%). As previously, no B. canis-positive ticks were found in Western Poland, including ticks from Wroclaw area (n = 298). DNA of B. canis was identified in 0.33% of juvenile ticks (in 3 pools of larvae and 2 nymphs) from the Eastern region. In the current study we confirmed high occurrence of R. raoultii in adults ticks from all four zones and relatively high prevalence of B. canis in the Eastern population of D. reticulatus, corresponding well with high incidence of canine babesiosis in this area of Poland. Finally, we confirmed R. raoultii and B. canis infection in all life stages of D. reticulatus ticks.

Immunolocalization of aquaporin-1, -5, and -7 in the avian testis and vas deferens.

Thirteen mammalian aquaporin (AQP) isoforms have been identified, and they have a unique tissue-specific pattern of expression. AQPs have been found in the reproductive system of both male and female humans, rats, and mice. However, tissue expression and cellular and subcellular localization of AQPs have been poorly investigated in the male reproductive system of birds. The localization of AQP subtypes (AQP1, 2, 3, 4, 5, 7, 8, 9, and 11) in the goose testis and vas deferens has been studied through immunohistochemistry and immunobloting. Interestingly, the testicular and deferential tissues were positive for AQP1, -5, and -7 but not the others. AQP1 immunoreactivity was detected in the capillary endothelial cells of testis and vas deferens. AQP5 was localized in the interstitial tissue of the testis, including Leydig cells, as well as in the basal cells of vas deferens. Double-labeling confocal microscopy revealed coexpression of AQP5 with capillary AQP1 in the testis. AQP7 was expressed in elongated spermatid and spermatozoa tails in the testis, as well as spermatozoa tails in the vas deferens. These results suggest that several subtypes of AQPs are involved in the regulation of water homeostasis in the goose male reproductive system.

Ontogeny of calcium-binding proteins in the cingulate cortex of the guinea pig: The same onset but different developmental patterns.

This paper compared the density of calbindin D28k (CB), calretinin (CR) and parvalbumin (PV) containing neurons in prenatal, newborn and postnatal periods in the cingulate cortex (CC) of the guinea pig as an animal model. The distribution and co-distribution among calcium-binding proteins (CaBPs) was also investigated during the entire ontogeny. The study found that CB-positive neurons exhibited the highest density in the developing CC. The CC development in the prenatal period took place with a high level of CB and CR immunoreactivity and both of these proteins reached peak density during fetal life. The density of PV-positive neurons, in contrast to CB and CR-positive neurons, reached high levels postnatally. The observed changes of the CaBPs-positive neuron density in the developing CC coincide with developmental events in the guinea pig. E.g. the eyes opening moment may be preceded by elevated levels of CB and CR at E50, whereas high immunoreactivity of PV from P10 to P40 with a peak at P20 may indicate the participation of PV in enhancement of the inhibitory cortical pathway maturation.

Calcium-binding proteins expression in the septum and cingulate cortex of the adult guinea pig.

For the first time this study demonstrates the distribution pattern and expression of three neuroanatomical markers: calbindin D28k (CB), calretinin (CR) and parvalbumin (PV) in topographically connected brain regions - the septum (SE) and the cingulate cortex (CC). The co-existence among calcium-binding proteins (CaBPs) was also examined. The study was conducted on the adult guinea pig with the use of immunohistochemical and molecular biological techniques. Among the studied CaBPs, which occurred in both examined brain regions at transcript and protein levels, CB was the most expressed in the SE, while CR in the CC. CR, unlike CB and PV, showed higher immunoreactivity in the superficial layers (II-III) of the CC than in the deep ones (V-VI). Most of CB and PV-positive perikarya were detected in the deep layers of the CC. Some CC neurons contained both CB and PV, suggesting cooperation between these CaBPs in the CC. Co-localization between CB and CR in the CC was not observed.

The ontogenetic development of neurons containing calcium-binding proteins in the septum of the guinea pig: Late onset of parvalbumin immunoreactivity versus calbindin and calretinin.

The study describes the immunoreactivity of calbindin (CB), calretinin (CR) and parvalbumin (PV), their distribution pattern and the co-distribution of CB and CR as well as CB and PV in the septum of the guinea pig during development. Immunohistochemistry was conducted on embryonic (E40, E50, E60), newborn (P0) and postnatal (P5, P10, P20, P40, P100) guinea pig brains. The presence of both CB and CR was detected at E40, while PV began to be observed at E60. Immunoreactivity for CB was constant throughout ontogeny. In contrast to CR immunoreactivity, PV immunoreactivity was higher in the postnatal stages than in the prenatal and newborn stages. Double immunostaining showed that CB co-localized with CR from E40 onwards, while with PV from P5 onwards, suggesting that CB co-operates with these proteins in the guinea pig septum during different periods of ontogeny. Our results also indicate that among the studied CaBPs, CB exhibited the highest immunoreactivity during both embryonic and postnatal development.

Distribution of cocaine- and amphetamine-regulated transcript (CART), neuropeptide Y (NPY) and galanin (GAL) in the pterygopalatine ganglion of the domestic duck (Anas platyrhynchos f. domestica).

INTRODUCTION: Cocaine- and amphetamine-regulated transcript (CART), neuropeptide Y (NPY) and galanin (GAL) act as neurotransmitters and neuromodulators in both the central and peripheral nervous systems. Their presence has been found in different taxonomic groups, in particular in mammals. However, only few investigators have studied these neuropeptides in the class Aves (birds). The aim of the present study was to describe the distribution of CART, NPY and GAL in the pterygopalatine ganglion (PPG) of the domestic duck (Anas platyrhynchos f. domestica). MATERIAL AND METHODS: The experiment was conducted on 16 one-year-old domestic ducks of the Pekin breed of both sexes (8 males and 8 females). Frozen sections of the PPG were subjected to immunofluorescence staining using primary mouse monoclonal antibodies directed against CART and GAL and rabbit polyclonal antibody directed against NPY. Secondary antibodies were conjugated with Cy3 and FITC fluorochromes. RESULTS: CART, NPY, and GAL were present in the PPG of the domestic duck. The highest immunoreactivity (IR) in the ganglionic cells was found for CART in the majority (83-85%) of neurons of both superior (SPPG) and inferior (IPPG) PPG. CART-IR was also found in small aggregations of neurons on the medial surface of the Harderian gland, and on the course of the palatine branch of the facial nerve. CART-IR was also observed in the nerve fibers of these neurons' aggregations; however, it was low in comparison to the immunoreactivity of the perikarya. Immunoreactivity of NPY was found in ganglionic neurons, but above all in numerous fibers of the SPPG and IPPG and within aggregations on the surface of the Harderian gland. NPY-IR cells were distributed irregularly over the cross-sections of the tested aggregations, and constituted from 36% to 43% of the SPPG and from 37% to 40% of the IPPG of all cross-sectioned neurons. GAL-immunoreactive perikarya, distributed irregularly across the sections, were observed in the SPPG, where they constituted 61-65%, and in the IPPG, where they made up 50-57% of all neurons. All immunoreactive neurons were characterized by immunopositive neuroplasm and immunonegative cell nuclei. CONCLUSIONS: The presence of CART, NPY, and GAL in the PPG of the domestic duck suggests that these peptides may contribute to the secretory innervation of the glands of the mucosa of the palate and nasal cavity, the Harderian gland, and the lacrimal gland.

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.

Tyrosine hydroxylase-immunoreactivity and its relations with gonadotropin-releasing hormone and neuropeptide Y in the preoptic area of the guinea pig.

The present study examines the distribution of tyrosine hydroxylase (TH) immunoreactivity and its morphological relationships with neuropeptide Y (NPY)- and gonadoliberin (GnRH)-immunoreactive (IR) structures in the preoptic area (POA) of the male guinea pig. Tyrosine hydroxylase was expressed in relatively small population of perikarya and they were mostly observed in the periventricular preoptic nucleus and medial preoptic area. The tyrosine hydroxylase-immunoreactive (TH-IR) fibers were dispersed troughout the whole POA. The highest density of these fibers was observed in the median preoptic nucleus, however, in the periventricular preoptic nucleus and medial preoptic area they were only slightly less numerous. In the lateral preoptic area, the density of TH-IR fibers was moderate. Two morphological types of TH-IR fibers were distinguished: smooth and varicose. Double immunofluorescence staining showed that TH and GnRH overlapped in the guinea pig POA but they never coexisted in the same structures. TH-IR fibers often intersected with GnRH-IR structures and many of them touched the GnRH-IR perikarya or dendrites. NPY wchich was abundantly present in the POA only in fibers showed topographical proximity with TH-IR structures. Althoug TH-IR perikarya and fibers were often touched by NPY-IR fibers, colocalization of TH and NPY in the same structures was very rare. There was only a small population of fibers which contained both NPY and TH. In conclusion, the morphological evidence of contacts between TH- and GnRH-IR nerve structures may be the basis of catecholaminergic control of GnRH release in the preoptic area of the male guinea pig. Moreover, TH-IR neurons were conatcted by NPY-IR fibers and TH and NPY colocalized in some fibers, thus NPY may regulate catecholaminergic neurons in the POA.

A morphometric study of the amygdala in the guinea pig.

The characteristic features of guinea pig amygdala (CA), as shown by volumetric comparisons of the individual nuclei, are the poor development of the basolateral (BL) and lateral olfactory tract (NLOT) nuclei as well as the strong formation of the lateral (LA) and basomedial (BM) nuclei. The central (CE), cortical (CO) and medial (ME) nuclei also appear to be well represented in this species. All these features are even more pronounced when the total number of neurons in the nuclei referred to was taken into consideration. A comparison of the densities of neurons in the individual nuclei with the mean numerical density of cells in the guinea pig CA indicates that the densities of neurons in LA, BL, BM, CE and CO are significantly lower than the mean (p < 0.05), whereas in the ME and NLOT these values are significantly higher than the mean (p < 0.05). It is noteworthy, that the densities of the neurons in CE and CO do not differ statistically from each other (p > 0.05) and are significantly higher than the respective values in LA, BL and BM (p < 0.05). Furthermore, a similar division of the guinea pig CA may to some extent be made using the size parameters of the amygdaloid neurons as a marker. Interestingly, the large neurons populate organised CA areas like LA, BL and BM less densely, whereas the small cells create ME and NLOT, where the neurons are densely arranged. CE and CO occupy intermediate positions, with the neurons similar in size to the mean for the guinea pig CA.

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.

The morphological types of neurones of the medial and lateral mamillary nuclei in a newborn guinea pig: Nissl, Klüver-Barrera and Golgi studies.

The studies were carried out on the hypothalamus of 5 newborn (P0 stage) guinea pigs. The sections were impregnated according to three modifications of the Golgi technique or stained according to the Nissl and Klüver-Barrera methods. On the basis of the shape and size of perikarya, dendroarchitecture, pattern of axon as well as the inner structure of neurones, in the medial (Mm) and lateral (MI) mamillary nuclei four morphological types of nerve cells were distinguished: cap-like with two subtypes (33% of the cell population), fusiform (35%), triangular (12%) and rounded unidendritic (21%) neurones. The majority of them possessed spines on their dendrites. The spiny cells, both cap-like and fusiform ones, were observed preponderantly, in the medial mamillary nucleus, whereas in the lateral mamillary nucleus there were mainly seen the triangular and fusiform neurones, either spiny or aspiny cells. The spineless rounded unidendritic cells were dispersed throughout the mamillary region, but they were twice as numerous in Mm as in MI, where they were the least numerous.

A morphometric study of the amygdala in the common shrew.

The characteristic features of the common shrew amygdala (CA), as shown by volumetric comparisons of the individual nuclei, are the poor development of the lateral (LA) and basomedial (BM) nuclei as well as the particularly strong formation of the basolateral (BL) and lateral olfactory tract (NLOT) nuclei. The central (CE), cortical (CO) and medial (ME) nuclei are also well organised in this species. All these features are even more distinctly visible when the total number of neurons in the nuclei referred to are compared. A comparison of the densities of neurons in the individual nuclei with the mean numerical density of cells in the CA indicates that there are the 3 different regions within the common shrew's CA. The densities of neurons in the LA, BL, and BM are significantly lower than the mean density of cells in the CA (p < 0.05). In the CE this value does not differ from the mean (p > 0.05). In the CO, ME and NLOT the density values are significantly higher than the mean (p < 0.05). Furthermore, a similar division of the shrew's CA can, to some extent, be performed using the size parameters of the amygdaloid neurons as a marker. Interestingly, the large neurons populate less densely organised CA areas like the LA, BL and BM, whereas the small cells populate the ME and NLOT, where the neurons are densely arranged. The CE and CO occupy intermediate positions, with the neurons similar in size to the mean for the shrew's CA.

A comparative study of the mammalian amygdala: a Golgi study of the basolateral amygdala.

The lateral (LA), basolateral (BL) and basomedial (BM) amygdaloid nuclei were compared in the guinea pig (Cavia porcellus), rabbit (Oryctolagus cuniculus) fox (Vulpes vulpes) and pig (Sus scrofa) by using the Golgi techniques. The interspecific comparisons of the individual basolateral nuclei have shown that the neuronal structure in each of them is extremely stable and remains almost unchanged in the series of animals studied. The only difference is the size of the basolateral neurons, which increases with the increasing size of the brain. Moreover, the intraspecific comparisons revealed that in all the animals studied LA, BL and BM form a fairly homogenous mass of cells in which similar cell types are present. The most numerous neurons in all basolateral nuclei are the spiny cells that often show a pyramidal or semi-pyramidal appearance (the Type I cells). Many of these have conical cell bodies and easily recognisable "apical" and "basal" dendrites. The Type II neurons are the most common variety of non-pyramidal cell and have round cell bodies and smooth or sparsely spined dendrites. The axons of these cells often form a dense terminal field that remains in the vicinity of the parent soma. The Type III cells, which are only occasionally seen, are small spine-sparse neurogliaform neurons with a few extremely delicate beaded dendrites and a poorly branching local axon. These neurons were only located in LA and BL.

The neuronal structure of the medial geniculate body in the pig--Nissi and Golgi study.

The studies were carried out on the brains of adult pigs. The preparations were made by means of the Golgi technique as well as the Nissl and Klüver-Barrera methods. Four types of neurons were described in the medial geniculate body (MGB) of the pig: 1. Multipolar neurons (perikarya 30-45 microm) with rounded, oval or quadrangular perikarya from which arise 4-7 dendritic trunks. The dendrites divide dichotomically twice, may send out collaterals and give off ramifications. The dendritic branches possess varicosities and knob-like spines. These neurons predominate in MGB. 2. Pear-shaped neurons (20-35 microm) with one or two dendritic trunks arising from one pole of the cell body. These dendrites have a tufted appearance. 3. Triangular neurons (30-45 microm) possess three thick dendrites which first bifurcate near the soma and then divide profusely into daughter branches. 4. Fusiform neurons (30-50 microm) have usually two dendritic trunks which arise from the opposite poles of the cell body and divide dichotomically twice. The fusiform neurons are the least numerous in MGB. Most MGB neurons have on the secondary tertiary dendrites and on their ramifications have delicate varicose or bead-like appendages and spine-like protrusions. In all types of neurons an axon arises either from the soma or from the initial portion of the dendritic trunk.

The neuronal structure of the red nucleus in newborn guinea pigs.

The preparations, stained according to the Nissl and Klüver-Barrera methods, were used to describe the topography and morphology of the red nucleus (RN) as well as the structure of the rubral perikarya in newborn (P0) guinea pigs. The Golgi impregnated preparations were used to distinguish types of neurons. RN is a uniform cell group and has the length from 740 to 860 microm. The Nissl stained perikarya were classified into three categories: big, medium-sized and small (A, B, C, respectively). The big perikarya, which contain a lot of tigroidal substance, were mainly observed at the caudal and ventral portions of RN. The small perikarya often have multiple nucleoli. The impregnated neurons were classified into 5 types: 1--large, aspiny, rich-arborised multipolar cells, 2--large and medium sized, spiny, rich-arborised fusiform or pear-shaped cells, 3--medium-sized, spiny, rich-arborised rounded cells, 4--medium-sized, spiny, rich-arborised bipolar cells, 5--small and single medium-sized cells. The 5th type constitutes a heterogeneous population and also has neurons in different developmental stages. Intraspecies variations concerning both the length of RN and a number of the triangular perikarya of the red nucleus were observed in the examined guinea pigs.

Calcium-binding proteins in the laterodorsal thalamic nucleus during development of the guinea pig.

The laterodorsal thalamic nucleus (LD) is often treated as a part of the anterior thalamic nuclei (ATN) because of its location and similar connectivity. Our previous studies have shown that distribution of three calcium-binding proteins, i.e. calbindin D28k (CB), calretinin (CR) and parvalbumin (PV), changes within the ATN during development of the guinea pig. The aim of this study is to examine the immunoreactivity pattern of these proteins in the LD in the guinea pig ontogeny. Brains from animals ranging from 40th embryonic day to 80th postnatal day were used in the study. Two methods were applied: a single-labelling immunoenzymatic method and double-labelling immunofluorescence. No changes of the distribution pattern of the substances were observed throughout the examined developmental stages. CB and CR were the most abundantly expressed proteins in perikarya of the LD. Numerous CB- and CR-immunoreactive cell bodies were found throughout the whole extent of the nucleus. In most of these cell bodies both proteins colocalized vastly. The highest immunoreactivity of the perikarya containing CB and CR was observed in the mediodorsal part of the LD and in its rostral portion. In regard to PV, single cell bodies were observed mostly in the dorsal part of the nucleus. PV did not colocalize with the other proteins. In summary, all the studied calcium-binding proteins were already present in the LD at prenatal developmental stages and the pattern of distribution remained virtually constant until adulthood. Thus, the LD differs considerably from the ATN in an aspect of neurochemical cell differentiation.

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.