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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Axotomy-induced changes in the chemical coding pattern of colon projecting calbindin-positive neurons in the inferior mesenteric ganglia of the pig.

The present study examines the response of colon-projecting neurons localized in the inferior mesenteric ganglia (IMG) to axotomy in the pig animal model. In all animals (n = 8), a median laparotomy was performed under anesthesia and the retrograde tracer Fast Blue was injected into the descending colon wall. In experimental animals (n = 4), the descending colon was exposed and the bilateral caudal colonic nerves were identified and severed. All animals were euthanized and the inferior mesenteric ganglia were harvested and processed for double-labeling immunofluorescence for calbindin-D28k (CB) in combination with either tyrosine hydroxylase (TH), neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), Leu-enkephalin (LENK), substance P, vesicular acetylcholine transporter, or galanin. Immunohistochemistry revealed significant changes in the chemical coding pattern of injured inferior mesenteric ganglion neurons. In control animals, Fast Blue-positive neurons were immunoreactive to TH, NPY, SOM, VIP, NOS, LENK, and CB. In the experimental group, the numbers of TH-, NPY-, and SOM-expressing neurons were reduced, whereas the number of neurons immunoreactive to LENK was increased. Our data indicate that the colon-projecting neurons of the porcine IMG react to the axotomy in a similar, but not an identical manner in a comparison to other species, especially rodents. Further studies are needed to elucidate the detailed factors/mechanisms involved in the response to nerve injury.

Developmental changes of calretinin immunoreactivity in the anterior thalamic nuclei of the guinea pig.

This study describes for the first time the distribution of the calcium-binding protein calretinin (CR) in the anterior thalamic nuclei (ATN) of the guinea pig during development. Brains from animals ranging from 40th embryonic day (E40) to 80th postnatal day (P80) were used in the study. No CR-immunoreactive (CR-ir) perikarya were present among the ATN at E40, but thick bundles of fibers containing CR were crossing the anteromedial nucleus (AM). The first CR-ir neurons appeared at E50 in the lateral part of the AM. At E60, the bundles of fibers disappeared and the whole area of AM displayed closely packed CR-ir perikarya. At this stage, CR also appeared in neurons of the anteroventral nucleus (AV), particularly in its lateral part and along its dorsal border. Moreover, from E50 short and thin bundles of fibers were observed in the medial part of the AV. The ATN of newborns (P0) already showed an adult-like CR distribution pattern - perikarya in the AM and AV were distributed more homogenously and their number was slightly decreased in comparison to E60. The anterodorsal nucleus (AD) was devoid of CR-ir neurons in all studied stages. In conclusion, our results demonstrate that calretinin appears for the first time in neurons of various anterior thalamic nuclei of the guinea pig between 40th and 60th day of prenatal development.

Proliferative enteropathy (PE)-induced changes in the calbindin-immunoreactive (CB-IR) neurons of inferior mesenteric ganglion supplying the descending colon in the pig.

A subpopulation of the pig inferior mesenteric ganglia (IMG) neurons projecting to the colon exhibit calbindin-like immunoreactivity. It is not known if there are any changes in the chemical coding patterns of these neurons during porcine proliferative enteropathy (PE). To answer this question, juvenile Large White Polish pigs with clinically diagnosed Lawsonia intracellularis infection (PE; n = 3) and a group of uninfected controls (C; n = 3) were compared. The retrograde tracer fast blue (FB) was injected into the descending colons of all animals and then tissue comprising IMGs from both groups was processed for double-labeling immunofluorescence with calbindin-D28k (CB) in combination with either tyrosine hydroxylase (TH), neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP), nitric oxide synthase, Leu-enkephalin, substance P, vesicular acetylcholine transporter, galanin, or pituitary adenylate cyclase-activating polypeptide. Immunohistochemistry revealed changes in the chemical coding pattern of calbindin-immunoreactive neurons in the inferior mesenteric ganglia of the pig. In control animals, FB/CB-positive neurons were immunoreactive to TH, NPY, SOM, and VIP. In the experimental group, TH-expressing neurons were unaffected, NPY-expressing neurons were increased, whereas the number of neurons immunoreactive to SOM or VIP was reduced. Changes in chemical coding of CB neurons during PE may play an important role in adaptation of these IMG cells under pathological conditions.

Inflammation-induced changes in the chemical coding pattern of colon-projecting neurons in the inferior mesenteric ganglia of the pig.

The present study examines the chemical coding of the inferior mesenteric ganglia after chemically induced colitis in the pig animal model. In all animals (n = 6), a median laparotomy was performed under anesthesia, and the Fast Blue retrograde tracer was injected into the descending colon wall. In experimental animals (n = 3), the thick descending colon were injected with formalin solution to induce inflammation. The animals were euthanized and the inferior mesenteric ganglion was harvested and processed for double-labeling immunofluorescence for calbindin-D28k (CB) in combination with either tyrosine hydroxylase (TH), neuropeptide Y (NPY), somatostatin (SOM), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), Leu-enkephalin (LENK), substance P (SP), vesicular acetylcholine transporter (VAChT), or galanin (GAL). Immunohistochemistry revealed significant changes in the chemical coding pattern of inferior mesenteric ganglion neurons. In control animals, Fast Blue-positive neurons were immunoreactive to TH, NPY, SOM, VIP, LENK, CB, and NOS. In the experimental group, TH, NPY, SOM, VIP, and LENK expressing neurons were reduced, whereas the number of neurons immunoreactive to CB, NOS, and GAL were increased. The increase of so-called neuroprotective neuropeptides suggests that the changes in the chemical coding of inferior mesenteric ganglion neurons reflect adaption under pathological conditions to promote their own survival.

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.

Distribution and chemical coding pattern of somatostatin immunoreactivity in the dorsal striatum of the guinea pig.

The present study provides a detailed description of somatostatin (SOM) distribution and the colocalization pattern of SOM, neuropeptide Y (NPY) and nitric oxide synthase (NOS) in the dorsal striatum (caudate-putamen complex) of the guinea pig. Within the dorsal striatum, SOM is found in a population of medium-sized aspiny interneurons. We found that 97% of all SOM-IR neurons expressed NPY simultaneously, while 98% of all NPY-ergic perikarya was simultaneously SOM-IR. On the other hand, while 98% of all SOM-IR cells were simultaneously NOS-IR, only 91% of all NOS-containing neurons exhibited SOM-immunoreactivity. Irrespective of their chemical coding, both types of SOM-IR neurons were scattered throughout the dorsal striatum, sometimes in the form of small, loosely arranged clusters of 2-4 cells. While SOM-IR and NPY-IR nerve fibers were present in all of the studied regions, they were more numerous in the ventro-medial part of the studied structure, with the exception of its caudal portion, where SOM-IR and NPY-IR fibers additionally formed a dense network in the part corresponding to the caudate nucleus. A low expression of staining for NOS-IR fibers was seen throughout the entire dorsal striatum. In some fibers, SOM and NPY were co-expressed. Fibers expressing both SOM and NOS were not found.